A novel structure-based approach for identification of vertebrate susceptibility to SARS-CoV-2: implications for future surveillance programmes

Understanding the origin of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been a highly debatable and unsolved challenge for the scientific communities across the world. A key to dissect the susceptibility profiles of animal species to SARS-CoV-2 is to understand how virus enters into the cells. The interaction of SARS-CoV-2 ligands (RBD on spike protein) with its host cell receptor, angiotensin-converting enzyme 2 (ACE2), is a critical determinant of host range and cross-species transmission. In this study, we developed and implemented a rigorous computational approach for predicting binding affinity between 299 ACE2 orthologs from diverse vertebrate species and the SARS-CoV-2 spike protein. The findings show that the spike protein of SARS-CoV-2 can bind to many vertebrate species carrying evolutionary divergent ACE2, implying a broad host range at the virus entry level, which may contribute to cross-species transmission and further viral evolution. Additionally, the present study facilitated the identification of genetic determinants that may differentiate susceptible from the resistant host species based on the conservation of ACE2- spike protein interacting residues in vertebrate host species known to facilitate SARS-CoV-2 infection; however, these genetic determinants warrant in vivo experimental confirmation. The molecular interactions associated with varied binding affinity of distinct ACE2 isoforms in a specific bat species were identified using protein structure analysis, implying the existence of diversified susceptibility of bat species to SARS-CoV-2. The findings from current study highlight the importance of intensive surveillance programs aimed at identifying susceptible hosts, particularly those with the potential to transmit zoonotic pathogens, in order to prevent future outbreaks.

First report of the zoonotic nematode Baylisascaris procyonis in non-native raccoons (Procyon lotor) from Italy

Baylisascaris procyonis is a nematode parasite of the raccoon (Procyon lotor), and it can be responsible for a severe form of larva migrans in humans. This parasite has been reported from many countries all over the world, after translocation of its natural host outside its native geographic range, North America. In the period between January and August 2021, 21 raccoons were cage-trapped and euthanized in Tuscany (Central Italy), in the context of a plan aimed at eradicating a reproductive population of this non-native species. All the animals were submitted for necroscopic examination. Adult ascariids were found in the small intestine of seven raccoons (prevalence 33.3%). Parasites have been identified as B. procyonis based on both morphometric and molecular approaches. The aim of the present article is to report the first finding of this zoonotic parasite from Italy, highlighting the sanitary risks linked to the introduction of alien vertebrate species in new areas. Graphical Abstract

Detection of Trypansoma cruzi in Kissing Bugs (Hemiptera: Reduviidae: Triatominae) Collected Across Oklahoma

Trypanosoma cruzi is the causative agent of Chagas disease in humans and dogs in the Americas. Transmission predominantly occurs via the feces of infected kissing bugs (Hemiptera: family Reduviidae; subfamily Triatominae) contaminating bite site wounds or mucous membranes. To better understand Chagas disease entomologic risk in Oklahoma, kissing bugs collected from within the state were tested for T. cruzi DNA. Data including county of insect collection, species and instar, and specific locations where specimens were found were collated. Triatomines were also tested by PCR to potentially identify DNA of vertebrate species on which specimens had recently fed. In total, 110 kissing bugs from 22 counties were tested. All triatomines were identified as Triatoma sanguisuga nymphs or adults, with the exception of one possible T. lecticularia adult. Trypanosoma cruzi DNA was detected in 22 (20%) triatomines from 12 counties spanning the state. The majority of T. cruzi PCR positive kissing bugs were found inside homes or associated structures (i.e., garages, porches). Vertebrate DNA was identified in 27 (24.5%) triatomines, with human DNA detected in 25 (92.6%) of these specimens, and canine and raccoon DNA detected in one specimen each (3.7%). Two specimens tested positive for both T. cruzi and human DNA and one specimen tested positive for both T. cruzi and raccoon DNA. Results from this study indicate that kissing bugs carrying T. cruzi are widespread in Oklahoma, that positive kissing bugs infest homes and associated structures, and that human-vector, canine-vector, and wildlife-vector contact all occur within the state.

Genoarchitectonic Compartmentalization of the Embryonic Telencephalon: Insights From the Domestic Cat

The telencephalon develops from the alar plate of the secondary prosencephalon and is subdivided into two distinct divisions, the pallium, which derives solely from prosomere hp1, and the subpallium which derives from both hp1 and hp2 prosomeres. In this first systematic analysis of the feline telencephalon genoarchitecture, we apply the prosomeric model to compare the expression of a battery of genes, including Tbr1, Tbr2, Pax6, Mash1, Dlx2, Nkx2-1, Lhx6, Lhx7, Lhx2, and Emx1, the orthologs of which alone or in combination, demarcate molecularly distinct territories in other species. We characterize, within the pallium and the subpallium, domains and subdomains topologically equivalent to those previously described in other vertebrate species and we show that the overall genoarchitectural map of the E26/27 feline brain is highly similar to that of the E13.5/E14 mouse. In addition, using the same approach at the earlier (E22/23 and E24/25) or later (E28/29 and E34/35) stages we further analyze neurogenesis, define the timing and duration of several developmental events, and compare our data with those from similar mouse studies; our results point to a complex pattern of heterochronies and show that, compared with the mouse, developmental events in the feline telencephalon span over extended periods suggesting that cats may provide a useful animal model to study brain patterning in ontogenesis and evolution.

Vertebrate Subfossil Localities in Taylorsville MetroPark, Montgomery County, Ohio, USA

Karst features in the Silurian dolomites of Taylorsville MetroPark (Dayton Metropolitan Area, Ohio, United States) were explored from 2017 to 2018 to identify sites of paleontological interest. Initial landscape surveys recovered 124 skeletal elements (from 12 sites) that were attributed to 17 vertebrate species—including evidence of such extirpated animals as bobcats (Lynx rufus) and rattlesnakes (Crotalus sp.). Of the 12 sites, 9 sites contained remains from the historical era and 3 sites contained much older remains (n = 17) that were radiocarbon dated to approximately 1,400 years before present (YBP). Human remains at one site, butchered bones at another, and artifacts from a third suggest a long period of pre-colonial human use of the area. The presence of rare taxa expands pre-historical species lists and confirms the coexistence of many previously undocumented taxa from the area during the late, pre-colonial, Holocene Epoch.

Comparative analysis of vertebrates reveals that mouse primordial oocytes do not contain a Balbiani body

Oocytes spend the majority of their lifetime in a primordial state. The cellular and molecular biology of primordial oocytes is largely unexplored; yet, studying these is necessary to understand the mechanisms through which oocytes maintain cellular fitness for decades, and why they eventually fail with age. Here, we develop enabling methods for live-imaging based comparative characterization of Xenopus, mouse and human primordial oocytes. We show that primordial oocytes in all three vertebrate species contain active mitochondria, Golgi apparatus and lysosomes. We further demonstrate that human and Xenopus oocytes have a Balbiani body characterized by a dense accumulation of mitochondria in their cytoplasm. However, despite previous reports, we did not find a Balbiani body in mouse oocytes. Instead, we demonstrate what was previously used as a marker for the Balbiani body in mouse primordial oocytes is in fact a ring-shaped Golgi apparatus that is not functionally associated with oocyte dormancy. Our work provides the first insights into the organisation of the cytoplasm in mammalian primordial oocytes, and clarifies relative advantages and limitations of choosing different model organisms for studying oocyte dormancy.

Genomic evidence for the parallel regression of melatonin synthesis and signaling pathways in placental mammals

Background: The study of regressive evolution has yielded a wealth of examples where the underlying genes bear molecular signatures of trait degradation, such as pseudogenization or deletion. Typically, it appears that such disrupted genes are limited to the function of the regressed trait, whereas pleiotropic genes tend to be maintained by natural selection to support their myriad purposes. One such set of pleiotropic genes is involved in the synthesis (AANAT, ASMT) and signaling (MTNR1A, MTNR1B) of melatonin, a hormone secreted by the vertebrate pineal gland. Melatonin provides a signal of environmental darkness, thereby influencing the circadian and circannual rhythmicity of numerous physiological traits. Therefore, the complete loss of a pineal gland and the underlying melatonin pathway genes seems likely to be maladaptive, unless compensated by extrapineal sources of melatonin. Methods: We examined AANAT, ASMT, MTNR1A and MTNR1B in 123 vertebrate species, including pineal-less placental mammals and crocodylians. We searched for inactivating mutations and modelled selective pressures (dN/dS) to test whether the genes remain functionally intact. Results: We report that crocodylians retain intact melatonin genes and express AANAT and ASMT in their eyes, whereas all four genes have been repeatedly inactivated in the pineal-less xenarthrans, pangolins, sirenians, and whales. Furthermore, colugos have lost these genes, and several lineages of subterranean mammals have partial melatonin pathway dysfunction. These results are supported by the presence of shared inactivating mutations across clades and analyses of selection pressure based on the ratio of non-synonymous to synonymous substitutions (dN/dS), suggesting extended periods of relaxed selection on these genes. Conclusions: The losses of melatonin synthesis and signaling date to tens of millions of years ago in several lineages of placental mammals, raising questions about the evolutionary resilience of pleiotropic genes, and the causes and consequences of losing melatonin pathways in these species.

Oxytocin has ‘Tend-and-Defend’ Functionality in Group Conflict Across Social Vertebrates

Across vertebrate species, intergroup conflict confronts individuals with a tension between group interests best served by participation in conflict and personal interest best served by not participating. Here, we identify the neurohormone oxytocin as pivotal to the neurobiological regulation of this tension in distinctly different group-living vertebrates, including fish, birds, rodents, non-human primates, and humans. In the context of intergroup conflict, a review of emerging work on pro-sociality suggests that oxytocin and its fish and birds homologs, isotocin and mesotocin, respectively, can elicit participation in group conflict and aggression. This is because it amplifies (i) concern for the interests of genetically related or culturally similar ‘in- group’ others, and (ii) willingness to defend against outside intruders and enemy conspecifics. Across a range of social vertebrates, oxytocin can induce aggressive behaviour to ‘tend-and- defend’ the in-group during intergroup contests.

A food web approach reveals the vulnerability of biocontrol services by birds and bats to landscape modification at regional scale

Pest control services provided by naturally occurring species (the so-called biocontrol services) are widely recognized to provide key incentives for biodiversity conservation. This is particularly relevant for vertebrate-mediated biocontrol services as many vertebrate species are of conservation concern, with most of their decline associated to landscape modification for agricultural purposes. Yet, we still lack rigorous approaches evaluating landscape-level correlates of biocontrol potential by vertebrates over broad spatial extents to better inform land-use and management decisions. We performed a spatially-explicit interaction-based assessment of potential biocontrol services in Portugal, using 1853 pairwise trophic interactions between 78 flying vertebrate species (birds and bats) and 53 insect pests associated to two widespread and economically valuable crops in the Euro-Mediterranean region, olive groves (Olea europaea subsp. europaea) and vineyards (Vitis vinifera subsp. vinifera). The study area was framed using 1004 square cells, each 10 × 10 km in size. Potential biocontrol services were determined at all those 10 × 10 km grid-cells in which each crop was present as the proportion of the realized out of all potential pairwise interactions between vertebrates and pests. Landscape correlates of biocontrol potential were also explored. Our work suggests that both birds and bats can effectively provide biocontrol services in olive groves and vineyards as they prey many insect pest species associated to both crops. Moreover, it demonstrates that these potential services are impacted by landscape-scale features and that this impact is consistent when evaluated over broad spatial extents. Thus, biocontrol potential by vertebrates significantly increases with increasing amount of natural area, while decreases with increasing area devoted to target crops, particularly olive groves. Overall, our study highlights the suitability of our interaction-based approach to perform spatially-explicit assessments of potential biocontrol services by vertebrates at local spatial scales and suggest its utility for integrating biodiversity and ecosystem services in conservation planning over broad spatial extents.