Infection agents of Didelphidae (Didelphimorphia) of Brazil: an underestimated matter in zoonoses research

Zoonoses are diseases or infections naturally transmissible from vertebrate animals to humans, and can be bacterial, viral or parasitic. The growth of urbanization, industrialization and the advance of agriculture and livestock facilitate the spread of infectious and parasitic agents from wild animals to the human population and to their domestic animals. Among the various reservoirs of zoonotic agents, we find that didelphid species, due to their high capacity for adaptation in urban environments, as an important study case. We reviewed the literature data on the pathogens, including with zoonotic potential of marsupial species occurring in Brazil, accounted for infections by agents that we categorized into Bacteria, Viruses, Protozoa, and Helminths. Aiming identifies possible knowledge gaps, we also surveyed the origin of studied samples and the institutions leading the researches on host didelphids. Among the hosts, the genus Didelphis in the cycles of these agents stands out. Moreover, we found that the majority of reported cases are in the Southeastern Brazil, mean the data from other Brazilian localities and didelphid species could be underestimated. Most studies took place in graduate programs of public research institutions, emphasizing the importance of the funding public research for the Brazilian scientific development.

Ancient Viral Integrations in Marsupials: A Potential Antiviral Defence

Marsupial viruses are understudied compared to their eutherian mammal counterparts, although they may pose severe threats to vulnerable marsupial populations. Genomic viral integrations, termed endogenous viral elements (EVEs) could protect the host from infection. It is widely known past viral infections and EVEs play an active role in antiviral defence in invertebrates and plants. This study aimed to characterise actively transcribed EVEs in Australian marsupial species, because they may play an integral role in cellular defence against viruses. This study screened publicly available RNA sequencing datasets (n=35) and characterised 200 viral transcripts from thirteen Australian marsupial species. Of the 200 transcripts, 188 originated from either Bornaviridae, Filoviridae or Parvoviridae EVEs. The other 12 transcripts were from putative active infections from members of the Herpesviridae and Anelloviridae, and Hepadnaviridae. EVE transcripts (n=188) were mapped to marsupial genomes (where available, n=5/13) to identify the genomic insertion sites. Of the 188 transcripts, 117 mapped to 39 EVEs within the koala, bare-nosed wombat, tammar wallaby, brushtail possum and Tasmanian devil genomes. The remaining eight animals had no available genome (transcripts n=71). Every marsupial have Bornaviridae, Filoviridae and Parvoviridae EVEs, a trend widely observed in eutherian mammals. Whilst eutherian bornavirus EVEs are predominantly nucleoprotein-derived, marsupial bornavirus EVEs demonstrate a surprising replicase gene bias. We predicted these widely distributed EVEs were conserved within marsupials from ancient germline integrations, as many were over 65 million years old. One bornavirus replicase EVE, present in six marsupial genomes, was estimated to be 160 million years old, predating the American-Australian marsupial split. We considered transcription of these EVEs through small non-coding RNA as an ancient viral defence. Consistent with this, in koala small RNA sequence datasets we detected Bornaviridae replicase and Filoviridae nucleoprotein produced piRNA. These were enriched in testis tissue, suggesting they could protect marsupials from vertically transmitted viral integrations.

Two Gaussian Bridge Processes for Mapping Continuous Trait Evolution along Phylogenetic Trees

Gaussian processes are powerful tools for modeling trait evolution along phylogenetic trees. As the value of a trait may change randomly throughout the evolution, two Gaussian bridge processes, the Brownian bridge (BB) and the Ornstein–Uhlenbeck bridge (OUB), are proposed for mapping continuous trait evolution for a group of related species along a phylogenetic tree, respectively. The corresponding traitgrams to the two bridge processes are created to display the evolutionary trajectories. The novel models are applied to study the body mass evolution of a group of marsupial species.

Detection of Cryptosporidium spp. and Giardia duodenalis in small wild mammals in northeastern Brazil

This study investigated the occurrence of Giardia duodenalis and Cryptosporidium spp. in rodents and marsupials from the Atlantic Forest in southern Bahia, northeastern Brazil. Two hundred and four fecal samples were collected from different forest areas in the municipalities of Ilhéus, Una, Belmonte, and Mascote. Identifications were performed using PCR and nested PCR followed by sequencing of the gdh and tpi genes for G. duodenalis, and the gp60 and Hsp-70 genes for Cryptosporidium. The total frequency of positive PCR samples for both G. duodenalis and Cryptosporidium spp. was 5.4% (11/204). Giardia duodenalis occurred in 2.94% (4/136) of rodents and 2.94% (2/68) of marsupials. The prevalence of Cryptosporidium in rodents and marsupials was 1.47% (2/136) and 4.41% (3/68), respectively. In the areas sampled, the frequency of parasitism was 50% (7/14), while the Mascote region alone had no parasitized animals. The G. duodenalis subgenotype AI was identified in the rodent species Hylaeamys laticeps, Oecomys catherinae, Oligoryzomys nigripes and Akodon cursor, and in the marsupials Gracilinanus agilis and Monodelphis americana. In the rodents Rhipidomys mastacalis, H. laticeps and in the marsupial Marmosa murina the protozoa Cryptosporidium fayeri, Cryptosporidium parvum and Cryptosporidium ubiquitum with subtypes IIa and IVg by the gp60 gene were found. In conclusion, this study provides the genetic characterization of Giardia and Cryptosporidium species and genotypes in rodents and marsupials. And, these findings reinforce that the rodent and marsupial species mentioned above play a role as new hosts for Giardia and Cryptosporidium.

Giardia and Cryptosporidium in Neo-Tropical Rodents and Marsupials: Is There Any Zoonotic Potential?

Cryptosporidiosis and giardiasis have been identified as emerging diseases in both developed and developing countries. Wildlife has been highlighted to play a major role in the spread of these diseases to humans. This review aims to highlight the research findings that relate to Cryptosporidium spp. and Giardia spp., with a focus on (1) parasitism of neo-tropical hystricomorphic rodents and marsupials from the genus Didelphis and (2) prevention and treatment strategies for humans and animals for the neo-tropical region. It was found that there are few studies conducted on neo-tropical rodent and marsupial species, but studies that were found illustrated the potential role these animals may play as zoonotic carriers of these two parasites for the neo-tropical region. Thus, it is recommended that further studies be done to assess the threat of protozoan parasites in neo-tropical wildlife to humans and domestic animals, and to further determine the most effective prophylaxis adapted for the unique conditions of the region.

Genome Sequences of Two Marsupial Simplex Viruses, Macropodid Alphaherpesviruses 2 and 4

ABSTRACT We present the genome sequences of macropodid alphaherpesviruses 2 and 4, two closely related pathogens of macropods. Both encoded 68 nonredundant open reading frames (ORFs) and share 90.6% genome-wide nucleotide identity. These viruses are associated with fatal outbreaks of disease in multiple marsupial species. These sequences will be important for the development of new diagnostic tools.

Viral fossils in marsupial genomes: secret cellular guardians

Genomic viral integrations, termed endogenous viral elements (EVEs), are fragments of viruses in host chromosomes that provide information about viral evolution and could even help protect the host from infection. In the present study we examined EVEs in thirteen different Australian marsupial species to identify trends in their integration, commonality and to investigate their possible cellular function. We found that marsupial EVEs are commonly derived from viruses of the Bornaviridae, Filoviridae and Parvoviridae families, and circulated up to 160 million years ago. We also show the EVEs are actively transcribed into both long and short RNA molecules in marsupials, and propose they are involved in a cellular defence mechanism to protect the germline from viral genomic invasion.

CD4, CD8b, and Cytokines Expression Profiles in Peripheral Blood Mononuclear Cells Infected with Different Subtypes of KoRV from Koalas (Phascolarctos cinereus) in a Japanese Zoo

Koala retrovirus (KoRV) poses a major threat to koala health and conservation, and currently has 10 identified subtypes: an endogenous subtype (KoRV-A) and nine exogenous subtypes (KoRV-B to KoRV-J). However, subtype-related variations in koala immune response to KoRV are uncharacterized. In this study, we investigated KoRV-related immunophenotypic changes in a captive koala population (Hirakawa zoo, Japan) with a range of subtype infection profiles (KoRV-A only vs. KoRV-A with KoRV-B and/or -C), based on qPCR measurements of CD4, CD8b, IL-6, IL-10 and IL-17A mRNA expression in unstimulated and concanavalin (Con)-A-stimulated peripheral blood mononuclear cells (PBMCs). Although CD4, CD8b, and IL-17A expression did not differ between KoRV subtype infection profiles, IL-6 expression was higher in koalas with exogenous infections (both KoRV-B and KoRV-C) than those with the endogenous subtype only. IL-10 expression did not significantly differ between subtype infection profiles but did show a marked increase—accompanying decreased CD4:CD8b ratio—in a koala with lymphoma and co-infected with KoRV-A and -B, thus suggesting immunosuppression. Taken together, the findings of this study provide insights into koala immune response to multiple KoRV subtypes, which can be exploited for the development of prophylactic and therapeutic interventions for this iconic marsupial species.