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To further understanding of the structure and morphology of the Orthohantavirus , family Hantaviridae , we have employed cryo-electron microscopy (cryo-EM) for three New World hantaviruses: Andes (ANDV), Sin Nombre (SNV), and Black Creek Canal (BCCV). Building upon our prior cryo-EM and cryo-tomography study of the Old World hantavirus, Hantaan virus (HTNV), we have expanded our studies to examine the entire virion population present in cell culture supernatant. Hence, in contrast to the prior cryo-EM/ET studies in which we used a polyethylene precipitation, a sucrose gradient, and a sucrose cushion, we used two sucrose cushions. We inactivated the material after the first cushion. We tested the method using HTNV which has a known cryo-EM structure and observed equivalent results. Therefore, we used this method to assess the particle distribution of the New World hantaviruses by cryo-EM. Cryo-EM images showed a diverse range of sizes and morphologies for the New World viruses that we classified as round, tubular, and irregular. Strikingly, BCCV virions were mostly tubular. These first cryo-EM images of the New World Orthohantavirus confirm prior EM observations that noted tubular projections of SNV at the plasma membrane during virion morphogenesis but were not confirmed. These findings underscore the need for further investigation of virion morphogenesis of the Orthohantavirus .

Begomoviruses, belonging to the family Geminiviridae and the genus Begomovirus, are DNA viruses that are transmitted by whitefly Bemisia tabaci (Gennadius) in a circulative persistent manner. They can easily adapt to new hosts and environments due to their wide host range and global distribution. However, the factors responsible for their adaptability and coevolutionary forces are yet to be explored. Among BGVs, TYLCV exhibits the broadest range of hosts. In this study, we have identified variable and coevolving amino acid sites in the proteins of Tomato yellow leaf curl virus (TYLCV) isolates from Old World (African, Indian, Japanese, and Oceania) and New World (Central and Southern America). We focused on mutations in the coat protein (CP), as it is highly variable and interacts with both vectors and host plants. Our observations indicate that some mutations were accumulating in Old World TYLCV isolates due to positive selection, with the S149N mutation being of particular interest. This mutation is associated with TYLCV isolates that have spread in Europe and Asia and is dominant in 78% of TYLCV isolates. On the other hand, the S149T mutation is restricted to isolates from Saudi Arabia. We further explored the implications of these amino acid changes through structural modeling. The results presented in this study suggest that certain hypervariable regions in the genome of TYLCV are conserved and may be important for adapting to different host environments. These regions could contribute to the mutational robustness of the virus, allowing it to persist in different host populations. [ABSTRACT FROM AUTHOR]

In this review, information was summarized on endoparasites found in six non-domesticated neotropical animals. These mammals have the potential to be domesticated. The animals included three rodents, agouti ( Dasyprocta leporina ), lappe ( Agouti paca ), and capybara ( Hydrochoerus hydrochaeris ); a marsupial, manicou ( Didelphis marsupialis insularis ); and an artiodactyl, the collared peccary ( Tayassu tajacu / Peccari tajacu ) and a ruminant (the red brocket deer, Mazama americana ). While there are many descriptions of the parasites present, the majority of publications failed to note the effect of them on the animals. Most information is available on endoparasites of capybara, while the endoparasites of the red brocket deer were the least reported. The manicou was reported to have had the most number of endoparasites, 44 species of parasites were reported, while there were only 24 endoparasites reported in the lappe. The most common parasites found in these neotropical animals were Paraspidodera uncinata , Strongyloides spp., Eimeria spp., Moniezia benedeni , Trichuris spp., Physocephalus spp., and Giardia spp. A large majority of the studies concluded that these animals were reservoirs for parasites that could affect domesticated livestock. Endoparasites of zoonotic significance were Echinoccocus spp., Trichuris spp., Giardia spp., and Cryptosporidium spp.

The begomoviruses (BGVs) are plant pathogens that evolved in the Old World during the Cretaceous and arrived to the New World (NW) in the Cenozoic era. A subgroup of NW BGVs, the " Squash leaf curl virus (SLCV) lineage" (S-Lin), includes viruses with unique characteristics. To get clues on the evolutionary origin of this lineage, a search for divergent members was undertaken. Four novel BGVs were characterized, including one that is basal to the group. Comparative analyses led to discover a ~670 bp genome module that is nearly exclusive of this lineage, encompassing the replication origin, the AC4 gene, and 480 bp of the Rep gene. A similar DNA module was found in two curtoviruses, hence suggesting that the S-Lin ancestor acquired its distinctive genomic segment by recombination with a curtovirus. This hypothesis was definitely disproved by an in-depth sequence analysis. The search for homologs of S-Lin Rep uncover the common origin of Rep proteins encoded by diverse Geminiviridae genera and viral "fossils" integrated at plant genomes. In contrast, no homolog of S-Lin Rep was found in public databases. Consequently, it was concluded that the SLCV clade ancestor evolved by a recombination event between a primitive NW BGV and a virus from a hitherto unknown lineage.

Mayaro (MAYV) and Una (UNAV) are emerging arboviruses belonging to the Alphavirus genus of the Togaviridae family. These viruses can produce febrile disease with symptoms such as fever, headache, myalgia, skin rash and incapacitating poly-arthralgia. Serological studies indicate that both viruses are circulating in different countries in Latin America. Viruses need the host cell machinery and resources to replicate effectively. One strategy to find new antivirals consists of identifying key cellular pathways or factors that are essential for virus replication. In this study, we analyzed the role of the ubiquitin-proteasome system (UPS) in MAYV and UNAV replication. Vero-E6 or HeLa cells were treated with the proteasome inhibitors MG132 or Lactacystin, and viral progeny production was quantified using a plaque assay method. In addition, the synthesis of viral proteins was analyzed by Western blot and confocal microscopy. Our results indicate that treatment with proteasome inhibitors decreases MAYV and UNAV protein synthesis, and also causes a significant dose-dependent decrease in MAYV and UNAV replication. Proteasome activity seems to be important at the early stages of MAYV replication. These findings suggest that the ubiquitin-proteasome system is a possible pharmacological target to inhibit these neglected alphaviruses.

Begomoviruses are one of the major groups of plant viruses with an important economic impact on crop production in tropical and subtropical regions. The global spread of its polyphagous vector, the whitefly Bemisia tabaci , has contributed to the emergence and diversification of species within this genus. In this study, we found a putative novel begomovirus infecting tomato plants in Venezuela without a cognate DNA-B component. This begomovirus was genetically characterized and compared with related species. Furthermore, its infectivity was demonstrated by agroinoculation of infectious clones in tomato ( Solanum lycopersicum ) and Nicotiana benthamiana plants. The name Tomato twisted leaf virus (ToTLV) is proposed. ToTLV showed the typical genome organization of the DNA-A component of New World bipartite begomoviruses. However, the single DNA component of ToTLV was able to develop systemic infection in tomato and N. benthamiana plants, suggesting a monopartite nature of its genome. Interestingly, an additional open reading frame ORF was observed in ToTLV encompassing the intergenic region and the coat protein gene, which is not present in other closely related begomoviruses. A putative transcript from this region was amplified by strand-specific reverse transcription-PCR. Along with recent studies, our results showed that the diversity of monopartite begomoviruses from the New World is greater than previously thought.

This review serves to summarize parasites found in Domesticated animals which were found in the Neo-Tropics. Indigenous domesticated Neo-tropical animals include South American camelids, ( Lama gunacoa , Lama glama , Lama pacos , Vicuna vicuna ), guinea pigs ( Cavia porcellus ), chinchillas ( Chinchilla lanigera ), turkeys ( Meleagris gallopavo ) and ducks ( Cairina moschata , Anas platyrhynchos , Dendrocyga autumnalis ). These animals were chosen due to their origin of existence (Neo-tropics) and over time these animals became domesticated and were distributed throughout the world. Over eighty (80) references were collected for this review and the papers spanned over eighty (80) years from 1934 to 2018. The gastrointestinal parasites reported for each animal were tabulated and their effects in the animal noted. Parasites reported in domesticated Neo-tropical animals had little to no effect on wild and free ranging animals with a few cases of illness and decreased productivity. The majority of articles viewed these animals as reservoir host which can infect humans and other domesticated livestock. It must also be noted that research done in the past did not focus on the effect these parasites had on these animals but only observed their potential as reservoirs for parasitic diseases.

The digital revolution has launched myriad new technologies in the field of art and cultural heritage law, including digital art, NFTs (non-fungible tokens), artificial intelligence (AI)-generated art, virtual reality and reality augmentation, online viewing rooms and auctions, holograms, immersive experiences, and more. As a $67.8 billion industry, the art market is a global driver of innovation, international collaboration, and national economies, given its cross-border transactions. However, given the extremely rapid development of these new technologies, regulators have struggled to keep pace and implement legal measures that are fit for purpose in this field. Limited oversight has resulted in several claims that have the potential to change the legal landscape. For instance, claims over the theft/misappropriation of NFTs and the related fraud and money laundering that may ensue, as well as a recent class action copyright infringement suit against the creators of a popular AI algorithm and infringement claims over immersive installation and light technologies, demonstrate how new ways of thinking are required to assess cases involving digital property (distinguished from other types of non-tangible property). Moreover, the US Supreme Court has issued a landmark ruling on fair use within the copyright context, which will be relied upon in the future to determine whether (and to what extent) the appropriation of existing copyrighted material is permitted. This includes both the digital use of physical artworks and the use of born-digital works. Although jurisprudential decisions are made on a case-by-case basis, factual patterns involving online media, digital art, and related technologies could serve as guidance for legislators and other decision-makers when considering what limits should be imposed on Web 3.0. This article will focus on recent US-based claims and regulations and dovetail with existing art market regulations in this jurisdiction (e.g., anti-money-laundering statutes) to determine their impact on new technologies, whether directly or indirectly. Finally, the article highlights ongoing trends and preoccupations to provide an overview of the shifting legal landscape. [ABSTRACT FROM AUTHOR]

The bifunctional enzyme Dihydrofolate reductase-thymidylate synthase (DHFR-TS) plays a crucial role in the survival of the Leishmania parasite, as folates are essential cofactors for purine and pyrimidine nucleotide biosynthesis. However, DHFR inhibitors are largely ineffective in controlling trypanosomatid infections, largely due to the presence of Pteridine reductase 1 (PTR1). Therefore, the search for structures with dual inhibitory activity against PTR1/DHFR-TS is crucial in the development of new anti-Leishmania chemotherapies. In this research, using the Leishmania major DHFR-TS recombinant protein, enzymatic inhibitory assays were performed on four kauranes and two derivatives that had been previously tested against LmPTR1. The structure 302 (6.3 µM) and its derivative 302a (4.5 µM) showed the lowest IC50 values among the evaluated molecules. To evaluate the mechanism of action of these structures, molecular docking calculations and molecular dynamics simulations were performed using a DHFR-TS hybrid model. Results showed that hydrogen bond interactions are critical for the inhibitory activity against LmDHFR-TS, as well as the presence of the p-hydroxyl group of the phenylpropanoid moiety of 302a. Finally, additional computational studies were performed on DHFR-TS structures from Leishmania species that cause cutaneous and mucocutaneous leishmaniasis in the New World (L. braziliensis, L. panamensis, and L. amazonensis) to explore the targeting potential of these kauranes in these species. It was demonstrated that structures 302 and 302a are multi-Leishmania species compounds with dual DHFR-TS/PTR1 inhibitory activity. [ABSTRACT FROM AUTHOR]

New to archaeological studies is the field of paleohydrology characterized by the use of modern hydraulic engineering and computational fluid dynamics (CFD) analysis of ancient urban and agricultural water supply and distribution systems. Examples are presented in nine chapters of CFD investigations of old and new world archaeological sites (several of which are World Heritage sites) using FLOW-3D CFD software to bring forward new discoveries revealing the depth of ancient water engineers' knowledge and creativity not previously noted in the archaeological literature. As modern analysis methods reveal technical details of ancient water systems, equivalent ancient technologies exist that were used in the design and operation of ancient water systems, albeit in formats, texts, and origins yet to be discovered. The nine chapters to follow present brief summaries of ancient sites' water systems and the use of CFD and modern hydraulic engineering methods to discover the water engineering knowledge base used by ancient water engineers. Results are new revelations unknown in the current literature of ancient sites. Paleohydrology studies presented serve to add a further dimension to the history of ancient new and old world archaeological sites by bringing forward added details of water engineering projects accomplished by ancient engineers. [ABSTRACT FROM AUTHOR]

The introduction of Zika virus (ZIKV) to the Americas raised concern that the virus would spill back from human transmission, perpetuated by Aedes aegypti , into a sylvatic cycle maintained in wildlife and forest-living mosquitoes. In the Americas, Sabethes species are vectors of sylvatic yellow fever virus (YFV) and are therefore candidate vectors of a sylvatic ZIKV cycle. To test the potential of Sabethes cyaneus to transmit ZIKV, Sa. cyaneus and Ae. aegypti were fed on A129 mice one or two days post-infection (dpi) with a ZIKV isolate from Mexico. Sa. cyaneus were sampled at 3, 4, 5, 7, 14, and 21 days post-feeding (dpf) and Ae. aegypti were sampled at 14 and 21 dpf. ZIKV was quantified in mosquito bodies, legs, and saliva to measure infection, dissemination, and potential transmission, respectively. Of 69 Sa. cyaneus that fed, ZIKV was detected in only one, in all body compartments, at 21 dpf. In contrast, at 14 dpf 100% of 20 Ae. aegypti that fed on mice at 2 dpi were infected and 70% had virus in saliva. These data demonstrate that Sa. cyaneus is a competent vector for ZIKV, albeit much less competent than Ae. aegypti .

Lactose maldigesters report an increase in abdominal pain due to the consumption of milk containing a mixture of A1 and A2 β-casein as compared to milk containing only A2 β-casein. Gastric transit affects gastrointestinal symptoms and rapid transit has been associated with an increase in abdominal pain. We conducted a double-blinded, randomized, crossover trial in 10 lactose maldigesters. Subjects consumed each of the two types of milk: conventional milk containing 75% A1 β-casein and 25% A2 β-casein and A2 milk containing 100% A2 β-casein. Magnetic resonance images were acquired, and abdominal pain was rated and recorded at 0, 10, 30, 60 and 120 min after milk consumption. The volume of milk in the stomach was calculated using FSL software. The volume of milk in the stomach after consuming milk with 75% A1 β-casein and 25% A2 β-casein was significantly lower at 30 (p = 0.01), 60 (p = 0.002) and 120 (p < 0.001) minutes as compared to milk with 100% A2 β-casein in the 10 lactose maldigesters. The transit of New-World milk containing A1 and A2 β-casein was more rapid as compared to Old-World milk containing only A2 β-casein. This difference in transit may mediate symptoms of lactose intolerance. [ABSTRACT FROM AUTHOR]

Traditional conservation practices have primarily relied on maintaining biodiversity by preserving species and habitats in place. Many regions are experiencing unprecedented environmental conditions, shifts in species distribution and habitats, and high turnover in species composition, resulting in ecological transformation. Natural resource managers have lacked tools for identifying and selecting strategies to manage ecosystem transformation. A recently formalized decision support framework provides a way for managers to resist, accept, or direct (RAD) the trajectory of change. We begin by identifying how historical conservation practices are built into the RAD framework. Next, we describe how RAD can be used to implement climate change adaptation actions, using examples from the Mojave Desert to provide ecological context. Third, we discuss how the RAD framework can assist with the creation of conservation portfolios, facilitating the maintenance of overall biodiversity across a landscape. Preserving species assemblages in their current state, or restoring them to historical conditions, will not always be possible, and RAD allows for explicit deliberation about when and where to prioritize scarce resources. We conclude with a set of guidelines for conservation practitioners or managers moving forward. Although operating under an increasingly uncertain future is daunting, managers can utilize RAD to conserve biodiversity and effectively handle ecosystem transformation. [ABSTRACT FROM AUTHOR]

Some New World (NW) and Old World (OW) mammalian arenaviruses are emerging, zoonotic viruses that can cause lethal hemorrhagic fever (HF) infections in humans. While these are closely related RNA viruses, the infected hosts appear to mount different types of immune responses against them. Lassa virus (LASV) infection, for example, results in suppressed immune function in progressive disease stage, whereas patients infected with Junín virus (JUNV) develop overt pro-inflammatory cytokine production. These viruses have also evolved different molecular strategies to evade host immune recognition and activation. This paper summarizes current progress in understanding the differential immune responses to pathogenic arenaviruses and how the information can be exploited toward the development of vaccines against them.

Concern about zoonoses and wildlife has increased. Few studies described the role of wild mammals and environments in the epidemiology of Salmonella. Antimicrobial resistance is a growing problem associated with Salmonella that threatens global health, food security, the economy, and development in the 21st century. The aim of this study is to estimate the prevalence and identify antibiotic susceptibility profiles and serotypes of non-typhoidal Salmonella enterica recovered from non-human primate feces, feed offered, and surfaces in wildlife centers in Costa Rica. A total of 180 fecal samples, 133 environmental, and 43 feed samples from 10 wildlife centers were evaluated. We recovered Salmonella from 13.9% of feces samples, 11.3% of environmental, and 2.3% of feed samples. Non-susceptibility profiles included six isolates from feces (14.6%): four non-susceptible isolates (9.8%) to ciprofloxacin, one (2.4%) to nitrofurantoin, and one to both ciprofloxacin and nitrofurantoin (2.4%). Regarding the environmental samples, one profile was non-susceptible to ciprofloxacin (2.4%) and two to nitrofurantoin (4.8%). The serotypes identified included Typhimurium/I4,[5],12:i:-, S. Braenderup/Ohio, S. Newport, S. Anatum/Saintpaul, and S. Westhampton. The epidemiological surveillance of Salmonella and antimicrobial resistance can serve in the creation of strategies for the prevention of the disease and its dissemination throughout the One Health approach. [ABSTRACT FROM AUTHOR]

Central and South American pitvipers, belonging to the genera Bothrops and Bothriechis, have independently evolved arboreal tendencies. Little is known regarding the composition and activity of their venoms. In order to close this knowledge gap, venom proteomics and toxin activity of species of Bothriechis, and Bothrops (including Bothriopsis) were investigated through established analytical methods. A combination of proteomics and bioactivity techniques was used to demonstrate a similar diversification of venom composition between large and small species within Bothriechis and Bothriopsis. Increasing our understanding of the evolution of complex venom cocktails may facilitate future biodiscoveries.

Multiple sclerosis (MS) is a highly heterogeneous disease involving a combination of inflammation, demyelination, and CNS injury. It is the leading cause of non-traumatic neurological disability in younger people. There is no cure, but treatments in the form of immunomodulatory drugs (IMDs) are available. Experience over the last 30 years has shown that IMDs, also sometimes called disease-modifying therapies, are effective in downregulating neuroinflammatory activity. However, there are a number of negatives in IMD therapy, including potential for significant side-effects and adverse events, uncertainty about long-term benefits regarding disability outcomes, and very high and increasing financial costs. The two dozen currently available FDA-approved IMDs also are heterogeneous with respect to efficacy and safety, especially long-term safety, and determining an IMD treatment strategy is therefore challenging for the clinician. Decisions about optimal therapy have been particularly difficult in early MS, at the time of the initial clinical demyelinating event (ICDE), at a time when early, aggressive treatment would best be initiated on patients destined to have a highly inflammatory course. However, given the fact that the majority of ICDE patients have a more benign course, aggressive immunosuppression, with its attendant risks, should not be administered to this group, and should only be reserved for patients with a more neuroinflammatory course, a decision that can only be made in retrospect, months to years after the ICDE. This quandary of moderate vs. aggressive therapy facing clinicians would best be resolved by the use of biomarkers that are predictive of future neuroinflammation. Unfortunately, biomarkers, especially molecular biomarkers, have not thus far been particularly useful in assisting clinicians in predicting the likelihood of future neuroinflammation, and thus guiding therapy. However, the last decade has seen the emergence of two highly promising molecular biomarkers to guide therapy in early MS: the CXCL13 index and neurofilament light. This paper will review the immunological and neuroscientific underpinnings of these biomarkers and the data supporting their use in early MS and will propose how they will likely be used to maximize benefit and minimize risk of IMDs in MS patients. [ABSTRACT FROM AUTHOR]

The New World Screwworm, Cochliomyia hominivorax (Coquerel), is one of the most damaging parasites of livestock, causing millions of dollars in annual losses to producers. The fly is an obligate parasite of warm-blooded animals, including humans. After a successful 50-year eradication campaign, C. hominivorax has been eradicated from the USA, Mexico and Central America by an area-wide integrated pest management approach. Recently, Caribbean and South American countries have expressed an interest in this approach. Aiming to support forthcoming projects in these countries, this review describes the main technical components of past and ongoing AW-IPM campaigns against C. hominivorax. [ABSTRACT FROM AUTHOR]

The familiar places where faculty and students engage, collaborate, debate, interact and exchange viewpoints appear to have been improved by introducing digital technology. This study investigates the influence of e-learning opportunities on faculty engagement in Nigerian universities. Five hundred faculty members were surveyed across eight private universities in Nigeria using purposive and convenient sampling techniques. Only 431 copies of the questionnaire, representing 86.2% response rate, were analysed with Smart PLS 3.0. The results show that virtual learning platforms, digital databases, online short courses and webinar learning platforms significantly influenced teaching, research, administrative and community engagements. The study concludes that the faculty of various universities should leverage e-learning platforms to be more engaged. The study recommends the machinery needed by the faculty members of Nigerian universities during the COVID-19 lockdown that challenged the conventional practice. The study empirically contributes to strengthening the current teaching, research, partnership and collaboration trends for improved faculty engagement in the new-normal world of work. [ABSTRACT FROM AUTHOR]

Simple Summary: The New World screwworm (NWS) fly is a pest related to economic impacts in livestock breeding, mainly because females lay their eggs in wounds and natural cavities of warm-blooded vertebrates. A direct way to control this pest is the sterile insect technique (SIT), which is based on the release of sexually sterilized males to mate with wild females, resulting in the absence of new flies. The first step to the implementation of such program is to know how flies' populations are structured, that is, if they can freely mate in the field or, otherwise, if they are impeded of mate due to spatial or temporal barriers. Here we investigated genomic polymorphisms to infer temporal structure in a population of the NWS fly from Uruguay in two consecutive years (approximately 18 generations). We found that this time was enough for this population to change its genetic composition, but it was not sufficient to prevent free mate among individuals from one year to the next. We also found signal that some of the polymorphisms are related to the response of the population to insecticide chemical control. This approach can be used in the future to estimate spatial barriers to geographically isolated populations. The New World screwworm (NWS) fly, Cochliomyia hominivorax (Diptera: Calliphoridae), is an economically important ectoparasite currently distributed in South America and in the Caribbean basin. The successful eradication of this species in USA, Mexico and continental Central America was achieved by a control program based on the sterile insect technique (SIT). In order to implement a genetic control strategy over the NWS fly's current area of occurrence, first, it is necessary to understand the species dynamics and population structure. In order to address this objective, the spatial genetic structure of the NWS fly was previously reported in South America based on different genetic markers; however, to date, no study has investigated temporal changes in the genetic composition of its populations. In the current study, the temporal genetic structure of a NWS fly population from Uruguay was investigated through two consecutive samplings from the same locality over an interval of approximately 18 generations. The genetic structure was accessed with neutral and under selection SNPs obtained with genotyping-by-sequencing. The results gathered with these data were compared to estimates achieved with mitochondrial DNA sequences and eight microsatellite markers. Temporal changes in the genetic composition were revealed by all three molecular markers, which may be attributed to seasonal changes in the NWS fly's southern distribution. SNPs were employed for the first time for estimating the genetic structure in a NWS fly population; these results provide new clues and perspectives on its population genetic structure. This approach could have significant implications for the planning and implementation of management programs. [ABSTRACT FROM AUTHOR]

Glasses, in their different forms and compositions, have special properties that are not found in other materials. The combination of transparency and hardness at room temperature, combined with a suitable mechanical strength and excellent chemical durability, makes this material indispensable for many applications in different technological fields (as, for instance, the optical fibres which constitute the physical carrier for high-speed communication networks as well as the transducer for a wide range of high-performance sensors). For its part, ion-exchange from molten salts is a well-established, low-cost technology capable of modifying the chemical-physical properties of glass. The synergy between ion-exchange and glass has always been a happy marriage, from its ancient historical background for the realisation of wonderful artefacts, to the discovery of novel and fascinating solutions for modern technology (e.g., integrated optics). Getting inspiration from some hot topics related to the application context of this technique, the goal of this critical review is to show how ion-exchange in glass, far from being an obsolete process, can still have an important impact in everyday life, both at a merely commercial level as well as at that of frontier research. [ABSTRACT FROM AUTHOR]

A specialized and fine-tuned immune response of bats upon infection with viruses is believed to provide the basis for a "friendly" coexistence with these pathogens, which are often lethal for humans and other mammals. First insights into the immunity of bats suggest that bats have evolved to possess their own strategies to cope with viral infections. Yet, the molecular details for this innocuous coexistence remain poorly described and bat infection models are the key to unveiling these secrets. In Jamaican fruit bats (Artibeus jamaicensis) , a New World bat species, infection experiments with its (putative) natural viral pathogens Tacaribe virus (TCRV), rabies virus (RABV), and the bat influenza A virus (IAV) H18N11, have contributed to an accurate, though still incomplete, representation of the bat-imposed immunity. Surprisingly, though many aspects of their innate and adaptive immune responses differ from that of the human immune response, such as a contraction of the IFN locus and reduction in the number of immunoglobulin subclasses, variations could also be observed between Jamaican fruit bats and other bat species.

Simple Summary: In this review, we provide updates on Sin Nombre virus (SNV) and other New World hantaviruses. We discuss the history of SNV, its biology and relatedness to other emerging New World hantaviruses, its ecology, and its disease in humans. This review highlights an important emerging virus pathogen from North America. Sin Nombre virus (SNV) is an emerging virus that was first discovered in the Four Corners region of the United States in 1993. The virus causes a disease known as Hantavirus Pulmonary Syndrome (HPS), sometimes called Hantavirus Cardiopulmonary Syndrome (HCPS), a life-threatening illness named for the predominance of infection of pulmonary endothelial cells. SNV is one of several rodent-borne hantaviruses found in the western hemisphere with the capability of causing this disease. The primary reservoir of SNV is the deer mouse (Peromyscus maniculatus), and the virus is transmitted primarily through aerosolized rodent excreta and secreta. Here, we review the history of SNV emergence and its virus biology and relationship to other New World hantaviruses, disease, treatment, and prevention options. [ABSTRACT FROM AUTHOR]

Plumolepilius Barrios-Izás & Anderson, 2016 is a leaf litter weevil genus that inhabits montane broadleaf forests from southern Mexico to northern Panama. The genus consists of 27 species, 22 distributed in the Chiapas Highlands province (Mexican Transition Zone) and 5 found in the Pacific dominion (Neotropical region) in Costa Rica and Panamá. Here, we analyze the phylogenetic relationships of the species of Plumolepilius based on 20 external body characters and 9 characters from the genitalia. The first dichotomy of the cladogram separates two species from the Pacific dominion from the remaining species of Plumolepilius from the Chiapas Highlands province and three species restricted to the Pacific dominion. We hypothesize that redundant distributions in the taxon-area cladogram of the genus may be due to dispersal events, probably during the Pleistocene glaciations. [ABSTRACT FROM AUTHOR]

Simple Summary: Fasciolid flukes collected from sheep and cattle in Ecuador showed a high diversity in DNA sequences whose analyses indicated introductions from South America, European and North American countries. These results agree with the numerous livestock importations performed by Ecuador. Abnormally big-sized liver flukes were found in Ecuadorian sheep. The morphometric phenotypic CIAS study showed that its size maximum and mean very pronouncedly and significantly surpassed those of the Fasciola hepatica populations from South America and Spain and proved to be intermediate between standard F. hepatica and F. gigantica populations. Such a feature is only known in intermediate fasciolid forms in Old World areas where the two species and their specific lymnaeid snail vectors overlap. This argues about a past hybridization after F. gigantica importation from Pakistan and/or introduction of intermediate hybrids previously generated in USA. The lack of heterozygotic rDNA ITS positions differentiating the two species, and of introgressed fragments and heteroplasmic positions in mtDNA genes, indicate a post-hybridization period sufficiently long as for rDNA concerted evolution to complete homogenization and mtDNA to return to homoplasmy. The vector specificity filter due to Radix absence should act as a driving force in accelerating such lineage evolution. Public health implications are finally emphasized. Fascioliasis is a disease caused by Fasciola hepatica worldwide transmitted by lymnaeid snails mainly of the Galba/Fossaria group and F. gigantica restricted to parts of Africa and Asia and transmitted by Radix lymnaeids. Concern has recently risen regarding the high pathogenicity and human infection capacity of F. gigantica. Abnormally big-sized fasciolids were found infecting sheep in Ecuador, the only South American country where F. gigantica has been reported. Their phenotypic comparison with F. hepatica infecting sheep from Peru, Bolivia and Spain, and F. gigantica from Egypt and Vietnam demonstrated the Ecuadorian fasciolids to have size-linked parameters of F. gigantica. Genotyping of these big-sized fasciolids by rDNA ITS-2 and ITS-1 and mtDNA cox1 and nad1 and their comparison with other countries proved the big-sized fasciolids to belong to F. hepatica. Neither heterozygotic ITS position differentiated the two species, and no introgressed fragments and heteroplasmic positions in mtDNA were found. The haplotype diversity indicates introductions mainly from other South American countries, Europe and North America. Big-sized fasciolids from Ecuador and USA are considered to be consequences of F.gigantica introductions by past livestock importations. The vector specificity filter due to Radix absence should act as driving force in the evolution in such lineages. [ABSTRACT FROM AUTHOR]

What is the connection between race and religious diversity? This question has emerged as particularly important in recent times, following heightened discussions on racial justice, equity, and the organization of society with regard to racial oppression. The terms race and religious diversity imply distinct points of contact that have within them diverse perspectives and worldviews, that carry with them assumed foundational understandings of the world and unexamined understandings of how the universe functions. This article explores the connection between race and religious diversity by discussing the physical and intellectual landscape and by raising concerns about the historical and religious-symbolic background of the Atlantic World. This background assumes the fact that the Atlantic World is more than just a historical phenomenon. Instead, the formation and operation of the Atlantic World reveals the construction of a cosmogony that informs racial (social/relational) and religious discourse (imagination/intellectual). The Atlantic World cosmogony arose from the conquest of the Americas by European interests, resulting in a terraforming process that adapted the New World to European sensibilities. The story of the Atlantic World cosmogony and the terraforming of the Americas serve as two points of reflection that call for assessing the connection of race and religious diversity. Concomitantly, considering the foreground of the Atlantic World cosmogony and terraforming opens the possibility of resituating the way we critically approach the discourse on race and religious diversity, allowing for communities to candidly express efforts to move beyond the history of the effects generated by the conquest of the Americas. [ABSTRACT FROM AUTHOR]

Iron oxide nanoparticles and single domain antibodies from camelids (VHHs) have been increasingly recognized for their potential uses for medical diagnosis and treatment. However, there have been relatively few detailed characterizations of their pharmacokinetics (PK). The aim of this study was to develop imaging methods and pharmacokinetic models to aid the future development of a novel family of brain MRI molecular contrast agents. An efficient near-infrared (NIR) imaging method was established to monitor VHH and VHH conjugated nanoparticle kinetics in mice using a hybrid approach: kinetics in blood were assessed by direct sampling, and kinetics in kidney, liver, and brain were assessed by serial in vivo NIR imaging. These studies were performed under "basal" circumstances in which the VHH constructs and VHH-conjugated nanoparticles do not substantially interact with targets nor cross the blood brain barrier. Using this approach, we constructed a five-compartment PK model that fits the data well for single VHHs, engineered VHH trimers, and iron oxide nanoparticles conjugated to VHH trimers. The establishment of the feasibility of these methods lays a foundation for future PK studies of candidate brain MRI molecular contrast agents.

Two major lineages of beetles inhabit cycad cones in the New World: weevils (Curculionoidea) in the subtribe Allocorynina, including the genera Notorhopalotria Tang and O’Brien, Parallocorynus Voss, Protocorynus O’Brien and Tang and Rhopalotria Chevrolat, and beetles in the family Erotylidae, including the genus Pharaxonotha Reitter. Analysis of the 16S ribosomal RNA (rRNA) mitochondrial gene as well as cladistic analysis of morphological characters of the weevils indicate four major radiations, with a probable origin on the cycad genus Dioon Lindl. and comparatively recent host shifts onto Zamia L. Analysis of the 16S rRNA gene for erotylid beetles indicates that an undescribed genus restricted to New World Ceratozamia Brongn. is the most early-diverging clade, and this lineage is sister to a large radiation of the genus Pharaxonotha onto Zamia, with apparent host shifts onto Dioon and Ceratozamia. Analysis of beetles are in accord with current models of continental drift in the Caribbean basin, support some proposed species groupings of cycads, but not others, and suggest that pollinator type may impact population genetic structure in their host cycads. [ABSTRACT FROM AUTHOR]

Rare species tend to be especially sensitive to habitat disturbance, making them important conservation targets. Thus, rarity patterns might be an important guide to conservation efforts. Rabinowitz's approach defines rarity using a combination of geographical range, habitat specificity, and local abundance, and is frequently used in conservation prioritization. Herein, we use Rabinowitz's approach to classify the New World (NW) pitvipers (family Viperidae) regarding rarity. We tested whether body size and latitude could predict rarity, and we compared rarity patterns with extinction risk assessments and other prioritization methods in order to detect rare species not classified as threatened or prioritized. Most NW pitvipers have large geographical ranges, high local abundances, and narrow habitat breadths. There are 11.8% of NW pitviper species in the rarest category and they occur along the Pacific coast of Mexico, in southern Central America, in the Andean region of Ecuador, and in eastern Brazil. Rarity in NW pitvipers is inversely related to latitude but is not related to body size. Our results indicate that additional species of NW pitvipers are threatened and/or should be prioritized for conservation. Combining complementary approaches to detect rare and threatened species may substantially improve our knowledge on the conservation needs of NW pitvipers. [ABSTRACT FROM AUTHOR]

The emergence of multiple concurrent infectious diseases localized in the world creates a complex burden on global public health systems. Outbreaks of Ebola, Lassa, and Marburg viruses in overlapping regions of central and West Africa and the co-circulation of Zika, Dengue, and Chikungunya viruses in areas with A. aegypti mosquitos highlight the need for a rapidly deployable, safe, and versatile vaccine platform readily available to respond. The DNA vaccine platform stands out as such an application. Here, we present proof-of-concept studies from mice, guinea pigs, and nonhuman primates for two multivalent DNA vaccines delivered using in vivo electroporation (EP) targeting mosquito-borne (MMBV) and hemorrhagic fever (MHFV) viruses. Immunization with MMBV or MHFV vaccines via intradermal EP delivery generated robust cellular and humoral immune responses against all target viral antigens in all species. MMBV vaccine generated antigen-specific binding antibodies and IFNγ-secreting lymphocytes detected in NHPs up to six months post final immunization, suggesting induction of long-term immune memory. Serum from MHFV vaccinated NHPs demonstrated neutralizing activity in Ebola, Lassa, and Marburg pseudovirus assays indicating the potential to offer protection. Together, these data strongly support and demonstrate the versatility of DNA vaccines as a multivalent vaccine development platform for emerging infectious diseases.

Small farm producers' sustenance depends on their alpaca herds and the production of fiber. Genetic improvement of fiber characteristics would increase their economic benefits and quality of life. The incorporation of molecular marker technology could overcome current limitations for the implementation of genetic improvement programs. Hence, the aim of this project was the generation of an alpaca single nucleotide polymorphism (SNP) microarray. A sample of 150 Huacaya alpacas from four farms, two each in Puno and Cerro de Pasco were used for SNP discovery by genotyping by sequencing (GBS). Reduced representation libraries, two per animal, were produced after DNA digestion with ApeK1 and double digestion with Pst1-Msp1. Ten alpaca genomes, sequenced at depths between 12× to 30×, and the VicPac3.1 reference genome were used for read alignments. Bioinformatics analysis discovered 76,508 SNPs included in the microarray. Candidate genes SNPs (302) for fiber quality and color are also included. The microarray SNPs cover 90.5% of the genome length with a density of about 39 ± 2.51 SNPs/Mb of DNA at an average interval of 26.45 ± 18.57 kbp. The performance was evaluated by genotyping 30 family trios and comparing them to their pedigrees, as well as comparing microarray to GBS genotypes. Concordance values of 0.93 and 0.94 for ApeK1 and Pst1-Msp1 generated SNPs were observed. Similarly, 290 fiber quality and color candidate gene SNPs were validated. Availability of this microarray will facilitate genome-wide association studies, marker-assisted selection and, in time, genomic selection.

The unique evolutionary dynamics and complex structure make the Y chromosome the most diverse and least understood region in the mammalian genome, despite its undisputable role in sex determination, development, and male fertility. Here we present the first contig-level annotated draft assembly for the alpaca ( Vicugna pacos ) Y chromosome based on hybrid assembly of short- and long-read sequence data of flow-sorted Y. The latter was also used for cDNA selection providing Y-enriched testis transcriptome for annotation. The final assembly of 8.22 Mb comprised 4.5 Mb of male specific Y (MSY) and 3.7 Mb of the pseudoautosomal region. In MSY, we annotated 15 X-degenerate genes and two novel transcripts, but no transposed sequences. Two MSY genes, HSFY and RBMY, are multicopy. The pseudoautosomal boundary is located between SHROOM2 and HSFY. Comparative analysis shows that the small and cytogenetically distinct alpaca Y shares most of MSY sequences with the larger dromedary and Bactrian camel Y chromosomes. Most of alpaca X-degenerate genes are also shared with other mammalian MSYs, though WWC3Y is Y-specific only in alpaca/camels and the horse. The partial alpaca Y assembly is a starting point for further expansion and will have applications in the study of camelid populations and male biology.

The Zika virus was introduced in Brazil in 2015 and, shortly after, spread all over the Americas. Nowadays, it remains present in more than 80 countries and represents a major threat due to some singularities among other flaviviruses. Due to its easy transmission, high percentage of silent cases, the severity of its associated complications, and the lack of prophylactic methods and effective treatments, it is essential to develop reliable and rapid diagnostic tests for early containment of the infection. Nonstructural protein 1 (NS1), a glycoprotein involved in all flavivirus infections, is secreted since the beginning of the infection into the blood stream and has proven to be a valuable biomarker for the early diagnosis of other flaviviral infections. Here, we describe the development of a highly sensitive nanobody ELISA for the detection of the NS1 protein in serum samples. Nanobodies were selected from a library generated from a llama immunized with Zika NS1 (ZVNS1) by a two-step high-throughput screening geared to identify the most sensitive and specific nanobody pairs. The assay was performed with a sub-ng/mL detection limit in the sera and showed excellent reproducibility and accuracy when validated with serum samples spiked with 0.80, 1.60, or 3.10 ng/mL of ZVNS1. Furthermore, the specificity of the developed ELISA was demonstrated using a panel of flavivirus' NS1 proteins; this is of extreme relevance in countries endemic for more than one flavivirus. Considering that the nanobody sequences are provided, the assay can be reproduced in any laboratory at low cost, which may help to strengthen the diagnostic capacity of the disease even in low-resource countries.

Botulinum neurotoxin (BoNT) serotype E is one of three serotypes that cause the preponderance of human botulism cases and is a Tier 1 Select Agent. BoNT/E is unusual among BoNT serotypes for its rapid onset and short duration of intoxication. Here we report two large panels of unique, unrelated camelid single-domain antibodies (VHHs) that were selected for their ability to bind to BoNT/E holotoxin and/or to the BoNT/E light chain protease domain (LC/E). The 19 VHHs which bind to BoNT/E were characterized for their subunit specificity and 8 VHHs displayed the ability to neutralize BoNT/E intoxication of neurons. Heterodimer antitoxins consisting of two BoNT/E-neutralizing VHHs, including one heterodimer designed using structural information for simultaneous binding, were shown to protect mice against co-administered toxin challenges of up to 500 MIPLD 50 . The 22 unique VHHs which bind to LC/E were characterized for their binding properties and 9 displayed the ability to inhibit LC/E protease activity. Surprisingly, VHHs selected on plastic-coated LC/E were virtually unable to recognize soluble or captured LC/E while VHHs selected on captured LC/E were poorly able to recognize LC/E coated to a plastic surface. This panel of anti-LC/E VHHs offer insight into BoNT/E function, and some may have value as components of therapeutic antidotes that reverse paralysis following BoNT/E exposures.

Coronaviruses are enveloped RNA viruses capable of causing respiratory, enteric, or systemic diseases in a variety of mammalian hosts that vary in clinical severity from subclinical to fatal. The host range and tissue tropism are largely determined by the coronaviral spike protein, which initiates cellular infection by promoting fusion of the viral and host cell membranes. Companion animal coronaviruses responsible for causing enteric infection include feline enteric coronavirus, ferret enteric coronavirus, canine enteric coronavirus, equine coronavirus, and alpaca enteric coronavirus, while canine respiratory coronavirus and alpaca respiratory coronavirus result in respiratory infection. Ferret systemic coronavirus and feline infectious peritonitis virus, a mutated feline enteric coronavirus, can lead to lethal immuno-inflammatory systemic disease. Recent human viral pandemics, including severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), and most recently, COVID-19, all thought to originate from bat coronaviruses, demonstrate the zoonotic potential of coronaviruses and their potential to have devastating impacts. A better understanding of the coronaviruses of companion animals, their capacity for cross-species transmission, and the sharing of genetic information may facilitate improved prevention and control strategies for future emerging zoonotic coronaviruses. This article reviews the clinical, epidemiologic, virologic, and pathologic characteristics of nine important coronaviruses of companion animals.

Background: Leishmaniasis, mainly cutaneous leishmaniasis (CL), is endemic in Israel. In recent years, the diagnosis of leishmaniasis has transitioned to a molecular diagnosis. Objective: To summarize all cases of leishmaniasis and the identified species seen in Israel based on molecular diagnosis. Methods: A retrospective study was performed of patients diagnosed with leishmaniasis between January 2017 and December 2022. All five medical centers in Israel in which Leishmania diagnosis is performed were included: Soroka, HaEmek, Hadassah, Rambam, and Sheba, all utilized molecular diagnostic methods. Data on the annual number of cases, species, age, and gender were retrieved. Results: During the years 2017–2022, a total of 4168 cases of leishmaniasis were diagnosed, which corresponds with ~7/100,000 inhabitants. L. (L.) major and L. (L.) tropica accounted for 84% and 14%, respectively. During the years 2020–2021, L. (L.) infantum emerged as a new form of cutaneous disease [2.7% of cases during this period]. Visceral L. (L.) infantum was found in five cases. Imported New World leishmaniasis accounted for 1% of the cases. L. (L.) major affected more males (67%) while L. (L.) tropica commonly affected more children and caused more facial lesions. Conclusions: The mean annual number of cases during these years is ~700. The dominant species is L. (L.) major. Since 2020, cutaneous L. (L.) infantum is an emerging infection in Israel. [ABSTRACT FROM AUTHOR]

Simple Summary: Thelyphonida, also known as vinegaroons or whip-scorpions, is a small arachnid order with 140 described species. Despite being conspicuous and widely distributed across the tropics and subtropics on four continents, knowledge of the order has been slow to advance. The genus Thelyphonellus, a member of subfamily Hypoctoninae, consisted of four species described prior to the present contribution. In this study, the first detailed morphological study and phylogenetic analysis of Thelyphonellus is presented. The analysis includes all except one of the previously described species of Thelyphonellus in addition to two new species described herein, the species of Ravilops (from the Caribbean island of Hispaniola), and the West African genus Etienneus. A single, optimal phylogenetic hypothesis placed Ravilops nested within Thelyphonellus. Four lineages with disjunct distributions, each characterized by a combination of characters, were recovered by the analysis of Hypoctoninae. Based on these results, four genera, two of which are new, are recognized: Ravilops, Thelyphonellus, Wounaan (new genus), and Yekuana (new genus). Two new species, classified in Wounaan and Yekuana, are described and illustrated. A key to the identification of the Neotropical genera of Hypoctoninae and a map plotting the known distribution of its species are also presented. Thelyphonida Blanchard, 1852, also known as vinegaroons or whip-scorpions, is a small arachnid order with 140 described species contained in a single family, Thelyphonidae Lucas, 1835. Despite being conspicuous and widely distributed across the tropics and subtropics on four continents, knowledge of the order has been slow to advance. Hypoctoninae Pocock, 1899, one of four subfamilies currently recognized and one of two represented in the New World, comprises five genera. Since its inception, Thelyphonellus Pocock, 1894 has remained the only hypoctonine genus occurring in South America, with only four species described prior to the present contribution. The first detailed morphological study and phylogenetic analysis of Thelyphonellus is presented herein. The morphological phylogenetic analysis—the first for Thelyphonida—includes all except one of the previously described species of Thelyphonellus in addition to two new species described herein; the species of Ravilops Víquez and Armas, 2005 (from the Caribbean island of Hispaniola); and the monotypic Old World genus Etienneus Heurtault, 1984 (from West Africa) scored for 45 morphological characters. A single, most parsimonious phylogenetic hypothesis revealed that Thelyphonellus is paraphyletic with respect to Ravilops. The New World Hypoctoninae comprises four clades with disjunct distributions and well supported by a combination of morphological characteristics, on the basis of which four genera, two of which are new, are recognized: Ravilops, with two species, endemic to Hispaniola; Thelyphonellus, herein restricted to Thelyphonellus amazonicus (Butler, 1872) and Thelyphonellus ruschii Weygoldt, 1979, occurring in Guyana, Suriname, French Guiana, and northern Brazil; Wounaan, gen. n., containing Wounaan vanegasae (Giupponi and Vasconcelos, 2008), comb. n. and Wounaan yarigui, sp. n. from Colombia; and Yekuana, gen. n., containing Yekuana venezolensis (Haupt, 2009), comb. n. and Yekuana wanadi, sp. n. from Venezuela. The two new species are described and illustrated. A key to the identification of the Neotropical genera of Hypoctoninae and a map plotting the known distribution of its species are also presented. [ABSTRACT FROM AUTHOR]

Llamas ( Lama glama ) are invaluable resources of Peru. Despite their importance, their population is decreasing. The Camelid Germplasm Bank-Quimsachata was created as a guardian of this South American camelid (SAC) species and established a bank of llamas from their two types, Ch'aku and Q'ara. However, these populations need to present high genetic diversity to be considered suitable conservation stocks. Thus, in the present study, 13 microsatellites specific for the SAC were used to assess the current genetic variability and differentiation of the llama population from the Bank. The global population showed high genetic diversity with a total of 157 different alleles, with an average of 12.08 alleles per microsatellite, an expected and observed heterozygosity of 0.758 and 0.707, respectively, and an average polymorphic information content (PIC) of 0.723. Although considered as two different breeds and managed separately, the genetic differentiation between Ch'aku and Q'ara was low (F ST = 0.01). Accordingly, the gene flow value was high (Nm = 30.5). Overall, our results indicate the existence of high genetic variation among individuals, and thus, this llama population could be considered a suitable genetic stock for their conservation and for sustainability programs. Additionally, the 13 microsatellites can be used to study other Peruvian llama populations and monitor the genetic variability of llamas from the Camelid Germplasm Bank-Quimsachata.

Alpaca is a camelid species of broad economic, biological and biomedical interest, and an essential part of the cultural and historical heritage of Peru. Recently, efforts have been made to improve knowledge of the alpaca genome, and its genetics and cytogenetics, to develop molecular tools for selection and breeding. Here, we report cytogenetic mapping of 35 new markers to 19 alpaca autosomes and the X chromosome. Twenty-eight markers represent alpaca SNPs, of which 17 are located inside or near protein-coding genes, two are in ncRNA genes and nine are intergenic. The remaining seven markers correspond to candidate genes for fiber characteristics ( BMP4, COL1A2, GLI1, SFRP4) , coat color ( TYR ) and development ( CHD7, PAX7 ). The results take the tally of cytogenetically mapped markers in alpaca to 281, covering all 36 autosomes and the sex chromosomes. The new map assignments overall agree with human-camelid conserved synteny data, except for mapping BMP4 to VPA3, suggesting a hitherto unknown homology with HSA14. The findings validate, refine and correct the current alpaca assembly VicPac3.1 by anchoring unassigned sequence scaffolds, and ordering and orienting assigned scaffolds. The study contributes to the improvement in the alpaca reference genome and advances camelid molecular cytogenetics.

Anti-idiotypic nanobodies, usually expressed by gene engineering protocol, has been shown as a nontoxic coating antigen for toxic compound immunoassays. We here focused on how to increase immunoassay sensitivity by changing the nanobody's primary sequence. In the experiments, two anti-idiotype nanobodies against monoclonal antibody 1H2, which is specific to ochratoxin A, were obtained and named as nontoxic coating antigen 1 (NCA1) and nontoxic coating antigen 2 (NCA2). Three differences between the nanobodies were discovered. First, there are six amino acid residues (AAR) of changes in the complementarity determining region (CDR), which compose the antigen-binding site. One of them locates in CDR1 (I-L), two of them in CDR2 (G-D, E-K), and three of them in CDR3 (Y-H, Y-W). Second, the affinity constant of NCA1 was tested as 1.20 × 10 8 L mol -1 , which is about 4 times lower than that of NCA2 (5.36 × 10 8 L mol -1 ). Third, the sensitivity (50% inhibition concentration) of NCA1 for OTA was shown as 0.052 ng mL -1 , which was 3.5 times lower than that of nontoxic coating antigen 2 (0.015 ng mL -1 ). The results indicate that the AAR changes in CDR of the anti-idiotypic nanobodies, from nonpolar to polar, increasing the affinity constant may enhance the immunoassay sensitivity. In addition, by using the nontoxic coating antigen 2 to substitute the routine synthetic toxic antigen, we established an eco-friendly and green enzyme-linked immunosorbent assay (ELISA) method for rapid detection of ochratoxin A in cereals. The half-maximal inhibitory concentration (IC 50 ) of optimized ELISA was 0.017 ng mL -1 with a limit of detection (LOD) of 0.003 ng mL -1 . The optimized immunoassay showed that the average recoveries of spiked corn, rice, and wheat were between 80% and 114.8%, with the relative standard deviation (RSD) ranging from 3.1-12.3%. Therefore, we provided not only basic knowledge on how to improve the structure of anti-idiotype nanobody for increasing assay sensitivity, but also an available eco-friendly ELISA for ochratoxin A in cereals.

In Europe, cowpox virus (CPXV) infection in South American camelids occurs as a so-called spill-over infection. Although infected animals generally have a mild form of the disease and survive, cases of fatal generalised CPXV infection have also been described. Prevention by prophylactic vaccination is the only way to protect animals from disease. In the present study, modified vaccinia virus Ankara (MVA) vaccine, which has been successfully used in many animal species, was used in a prime-boost vaccination regimen in two alpaca herds with a history of CPXV infection. The focus of the study was the prevention of further clinical cases, and to determine the safety and immunogenicity of the MVA vaccine in alpacas. The MVA vaccine was well tolerated and safe in the 94 animals vaccinated. An indirect immunofluorescence assay (IFA) using MVA as an antigen showed that the seroprevalence of antibody after booster vaccination was 81.3% in herd I and 91.7% in herd II. Detectable antibody titres declined to 15.6% in herd I and 45.8% in herd II over a 12-month period after booster vaccination. Animals could be divided into four groups based on individual antibody titres determined over one year: Group 1 consisted of 19.3% of animals that were seropositive until the end of the trial period; Group 2 consisted of 58.0% of animals that were seropositive after booster vaccination, but seronegative one year later; Group 3 consisted of 14.7% of animals that were not seropositive at any time point; and Group 4 consisted of 7.9% of animals that were seropositive after initial immunisation, seronegative six months later, but seropositive or intermediate in IFA one year after immunisation, likely because of natural exposure. In new-born crias born to MVA-vaccinated mares, specific maternal antibodies were detected in 50.0% of animals up to 14 weeks of age. Our results confirm that MVA vaccination is a feasible tool for the prevention of CPXV disease in alpacas. Long-term studies are needed to verify future vaccination regimen in CPXV affected herds.

The emergence and re-emergence of porcine reproductive and respiratory syndrome virus (PRRSV) has resulted in huge economic losses for the swine industry. Current vaccines are of limited efficacy against endemic circulating PRRSV variants. New strategies against PRRSV infection are in urgent need. Here, a nanobody library in Marc-145 cells is constructed for antiviral nanobodies. Nanobody encoding sequences from two non-immunized llamas were cloned to generate a pseudotyped lentiviral library. Several candidates were selected from survival cells post-PRRSV inoculation and further characterized. Nb9 was identified with strong antiviral activity. Moreover, Nb9 exerted antiviral activity via its interaction with PRRSV viral proteins, as revealed by immunofluorescence assay and Western blot. Taken together, the novel function-based screen of the lentivirus nanobody library, instead of the conventional affinity-based screen, offers an alternative strategy for antiviral reagents against PRRSV and other pathogens.

Overexpression of human epidermal growth factor receptor type 2 (HER2) is considered as a prognostic factor of breast cancer, which is positively associated with recurrence when cancer metastasizes to the lymph nodes. Here, we expressed the single variable domain on a heavy chain (VHH) form of anti-HER2 camelid single domain antibody in tobacco plants and compared its in vitro anticancer activities with the anti-HER2 full size antibody. The gene expression cassette containing anti-HER2 camelid single domain antibody VHH fused to human IgG Fc region with KDEL endoplasmic reticulum (ER) (VHH-FcK) was transferred into the tobacco plant via the Agrobacterium -mediated transformation. The transformants were screened with polymerase chain reaction and Western blot analyses. Enzyme-linked immunosorbent assay (ELISA) confirmed the binding of the purified anti-HER2 VHH-FcK to the HER2-positive breast cancer cell line, SK-BR-3. Migration assay results confirmed anticancer activity of the plant-derived anticancer camelid single chain antibody. Taken together, we confirmed the possibility of using anti-HER2 VHH-FcK as a therapeutic anticancer agent, which can be expressed and assembled and purified from a plant expression system as an alternative antibody production system.

Monitoring the cardiopulmonary signal of animals is a challenge for veterinarians in conditions when contact with a conscious animal is inconvenient, difficult, damaging, distressing or dangerous to personnel or the animal subject. In this pilot study, we demonstrate a computer vision-based system and use examples of exotic, untamed species to demonstrate this means to extract the cardiopulmonary signal. Subject animals included the following species: Giant panda (Ailuropoda melanoleuca), African lions (Panthera leo), Sumatran tiger (Panthera tigris sumatrae), koala (Phascolarctos cinereus), red kangaroo (Macropus rufus), alpaca (Vicugna pacos), little blue penguin (Eudyptula minor), Sumatran orangutan (Pongo abelii) and Hamadryas baboon (Papio hamadryas). The study was done without need for restriction, fixation, contact or disruption of the daily routine of the subjects. The pilot system extracts the signal from the abdominal-thoracic region, where cardiopulmonary activity is most likely to be visible using image sequences captured by a digital camera. The results show motion on the body surface of the subjects that is characteristic of cardiopulmonary activity and is likely to be useful to estimate physiological parameters (pulse rate and breathing rate) of animals without any physical contact. The results of the study suggest that a fully controlled study against conventional physiological monitoring equipment is ethically warranted, which may lead to a novel approach to non-contact physiological monitoring and remotely sensed health assessment of animals. The method shows promise for applications in veterinary practice, conservation and game management, animal welfare and zoological and behavioral studies.

Nanobodies are one-tenth the size of conventional antibodies and are naturally obtained from the atypical heavy-chain-only antibodies present in camelids. Their small size, high solubility, high stability, and strong resilience to organic solvents facilitate their use as novel analytical reagents in immunochemistry. In this study, specific nanobodies against pesticide carbofuran were isolated and characterized from an immunized library via phage display platform. We further established an indirect competitive enzyme-linked immunosorbent assay (ELISA) using nanobody Nb316 to detect carbofuran in vegetable and fruit samples. The results showed a half-maximal inhibitory concentration (IC 50 ) of 7.27 ng/mL and a detection limit of 0.65 ng/mL. A simplified sample pretreatment procedure omitting the evaporation of organic solvent was used. The averaged recovery rate of spiked samples ranged between 82.3% and 103.9%, which correlated with that of standard UPLC-MS/MS method. In conclusion, a nanobody with high specificity for carbofuran was characterized, and a nanobody-based sensitive immunoassay for simple and rapid detection of carbofuran in real samples was validated.

The bacterium Clostridium botulinum is the causative agent of botulism-a severe intoxication caused by botulinum neurotoxin (BoNT) and characterized by damage to the nervous system. In an effort to develop novel C. botulinum immunotherapeutics, camelid single-domain antibodies (sdAbs, VHHs, or nanobodies) could be used due to their unique structure and characteristics. In this study, VHHs were produced using phage display technology. A total of 15 different monoclonal VHHs were selected based on their comlementarity-determining region 3 (CDR3) sequences. Different toxin lethal dose (LD 50 ) challenges with each selected phage clone were conducted in vivo to check their neutralizing potency. We demonstrated that modification of neutralizing VHHs with a human immunoglobulin G (IgG)1 Fc (fragment crystallizable) fragment (fusionbody, VHH-Fc) significantly increased the circulation time in the blood (up to 14 days). At the same time, VHH-Fc showed the protective activity 1000 times higher than monomeric form when challenged with 5 LD 50 . Moreover, VHH-Fcs remained protective even 14 days after antibody administration. These results indicate that this VHH-Fc could be used as an effective long term antitoxin protection against botulinum type A., Competing Interests: The authors declare no conflict of interest.

Viruses belonging to the genus Bocaparvovirus ( ) herd with reoccurring diarrhea and respiratory disease was submitted for next-generation sequencing, revealing the presence of a BoV strain. The alpaca BoV strain (AlBoV) had a 58.58% whole genome nucleotide percent identity to a camel BoV from Dubai, belonging to a tentative ungulate BoV 8 species (UBoV8). Recombination events were lacking with other UBoV strains. The AlBoV genome was comprised of the NS1, NP1, and VP1 proteins. The NS1 protein had the highest amino acid percent identity range (57.89-67.85%) to the members of UBoV8, which was below the 85% cut-off set by the International Committee on Taxonomy of Viruses. The low NS1 amino acid identity suggests that AlBoV is a tentative new species. The whole genome, NS1, NP1, and VP1 phylogenetic trees illustrated distinct branching of AlBoV, sharing a common ancestor with UBoV8. Walker loop and Phospholipase A2 (PLA2) motifs that are vital for virus infectivity were identified in NS1 and VP1 proteins, respectively. Our study reports a novel BoV strain in an alpaca intestinal sample and highlights the need for additional BoV research.BoV ) are a genetically diverse group of DNA viruses known to cause respiratory, enteric, and neurological diseases in animals, including humans. An intestinal sample from an alpaca ( Vicugna pacos ) herd with reoccurring diarrhea and respiratory disease was submitted for next-generation sequencing, revealing the presence of a BoV strain. The alpaca BoV strain (AlBoV) had a 58.58% whole genome nucleotide percent identity to a camel BoV from Dubai, belonging to a tentative ungulate BoV 8 species (UBoV8). Recombination events were lacking with other UBoV strains. The AlBoV genome was comprised of the NS1, NP1, and VP1 proteins. The NS1 protein had the highest amino acid percent identity range (57.89-67.85%) to the members of UBoV8, which was below the 85% cut-off set by the International Committee on Taxonomy of Viruses. The low NS1 amino acid identity suggests that AlBoV is a tentative new species. The whole genome, NS1, NP1, and VP1 phylogenetic trees illustrated distinct branching of AlBoV, sharing a common ancestor with UBoV8. Walker loop and Phospholipase A2 (PLA2) motifs that are vital for virus infectivity were identified in NS1 and VP1 proteins, respectively. Our study reports a novel BoV strain in an alpaca intestinal sample and highlights the need for additional BoV research., Competing Interests: The authors declare no conflict of interest

MERS-CoV is present in dromedary camels throughout the Middle East and Africa. Dromedary camels are the primary zoonotic reservoir for human infections. Interruption of the zoonotic transmission chain from camels to humans, therefore, may be an effective strategy to control the ongoing MERS-CoV outbreak. Here we show that vaccination with an adjuvanted MERS-CoV Spike protein subunit vaccine confers complete protection from MERS-CoV disease in alpaca and results in reduced and delayed viral shedding in the upper airways of dromedary camels. Protection in alpaca correlates with high serum neutralizing antibody titers. Lower titers of serum neutralizing antibodies correlate with delayed and significantly reduced shedding in the nasal turbinates of dromedary camels. Together, these data indicate that induction of robust neutralizing humoral immune responses by vaccination of naïve animals reduces shedding that potentially could diminish the risk of zoonotic transmission.

Indicators have been used in many sustainability assessment methods, however, disagreements over a common definition and scope for the sustainability concept have led to many distinct assessment methods, which are not often directly comparable. Before developing a sustainability assessment, it is essential to: define sustainability and specify the viewpoint of the assessor, the purpose of the assessment, and the context and time frame of the assessment. This article presents a short list of indicators and a method that can be readily adopted by any agricultural business or region to assess sustainability, making any organization, region or crop qualitatively comparable. These indicators were proposed by 83 top-level executives in 14 group interviews conducted using our adapted nominal group technique (ANGT). Executives were sourced from wine-grape growing organisations from New World wine-producing countries that also owned vineyards, and they considered everyday management practices of farms. These indicators, grouped within three categories (economic, environmental, and social) were ranked by their importance. The method defines qualitative indicators that, in the context of distinct wine regions or crops should be quantified to maintain their relevance and usefulness. [ABSTRACT FROM AUTHOR]