Search Results (15)

Over 40 years ago, scientists imagined ways cloning could aid conservation of threatened taxa. The cloning of Dolly the sheep from adult somatic cells in 1996 was the breakthrough that finally enabled the conservation potential of the technology. Until the 2020s, conservation cloning research efforts yielded no management applications, leading many to believe cloning is not yet an effective conservation tool. In strong contrast, domestic taxa are cloned routinely for scientific and commercial purposes. In this review, we sought to understand the reasons for these divergent trends. We scoured peer-reviewed and gray literature and sent direct inquiries to scientists to analyze a more comprehensive history of the field than was analyzed in previous reviews. While most previous reviewers concluded that a lack of reproductive knowledge of wildlife species has hindered advances for wider conservation applications, we found that resource limitations (e.g., numbers of surrogates, sustainable funding) and widely held misconceptions about cloning are significant contributors to the stagnation of the field. Recent successes in cloning programs for the endangered black-footed ferret (Mustela nigripes) and Przewalski’s horse (Equus przewalskii), the world’s first true applied-conservation cloning efforts, are demonstrating that cloning can be used for significant conservation impact in the present. When viewed alongside the long history of cloning achievements, these programs emphasize the value of investing in the science and resources needed to meaningfully integrate cloning into conservation management, especially for species with limited genetic diversity that rely on the maintenance of small populations for many generations while conservationists work to restore habitat and mitigate threats in the wild. Full article

The present study aimed to describe the reproductive cycle of the black-footed limpet (Patella depressa Pennant, 1777) from an intertidal rocky shore on the Algarve coast (southern Portugal). Samples were collected monthly between January 2017 and December 2018, with the species’ gametogenic cycle being described based on gonad histology and the mean gonadal index. The presence of both transitional and mosaic hermaphrodites indicates that some individuals are able to change sex (sequential hermaphroditism). Despite the occurrence of hermaphroditism, sex proportions were approximately equal, suggesting the absence of protandric sex change in this species. The population exhibited an extensive occurrence of ripe and spawning gonads throughout almost the whole study period, probably related to consecutive processes of gonadal re-ripening and partial spawning events. The reproductive dynamics of P. depressa displayed clear inter-annual differences, with a short resting period recorded in 2017 (June–August) and the absence of resting gonads in 2018. The continued monitoring of this population and collection of environmental data are required to further improve knowledge of the reproductive dynamics of this species. Such information is crucial for proposing additional management measures for the sustainable harvesting of limpets in southern Portugal. Full article

Arsenic contamination in groundwater poses serious health risks, as exemplified by the Blackfoot disease epidemic in Taiwan, which was caused by prolonged arsenic exposure. This study investigates the use of biochar derived from the wastewater treatment sludge of the Far Eastern Memorial Hospital (New Taipei City, Taiwan) as an efficient adsorbent for arsenic removal. A novel iron-doped sludge biochar (Fe-SBC) was developed to enhance arsenic adsorption efficiency, facilitate adsorbent recovery, and reduce operational costs. The adsorption mechanism of arsenic on Fe-SBC, modified with iron hydroxide complexes, was examined through Density Functional Theory (DFT) simulations. The results demonstrate a high arsenic removal efficiency of approximately 90% using continuous adsorption systems. The DFT calculations revealed strong chemical interactions between arsenic and the biochar, evidenced by high adsorption energy (−156.8 kJ mol⁻¹) and a short bond distance (1.48 Å), correlating with the high adsorption performance observed experimentally. Additionally, arsenic byproducts desorbed from the adsorbent were repurposed into antibacterial agents and pigments. Four distinct pigment colors—green, blue, gray, and orange—were produced through different preparation methods, with the antibacterial agents showing effective antimicrobial properties. This study highlights the potential of Fe-SBC for sustainable arsenic remediation and resource recovery. Full article

The black-footed ferret (Mustela nigripes), a North American mustelid species, was once found abundantly throughout the Midwest until the extreme decline in prairie dogs (Cynomys spp.), the black-footed ferret’s primary food source, brought the species to near-extinction. Subsequently, the Black-Footed Ferret Recovery Program was created in the 1980s with a goal of bringing all remaining individuals of the species into captivity in order to breed the species back to a sustainable population level for successful reintroduction into the wild. While many components of the ferrets’ health were accounted for while in captivity—especially those affecting fecundity—this study aims to assess the effects that captivity may have had on their cranial morphology, something that has not been widely studied in the species. In a previous study, we showed that the captive ferrets had significant oral health problems, and here we aim to document how the captive diet also affected their skull shape. For this study, 23 cranial measurements were taken on the skulls of 271 adult black-footed ferrets and 53 specimens of two closely related species. Skulls were divided based on sex, species, captivity status and phase of captivity and compared for all measurements using stepwise discriminant analysis as well as principal component analysis derived from the combined variables. We found that there are significant differences between captive and wild specimens, some of which are larger than interspecific variation, and that a diet change in the captive specimens likely helped decrease some of these differences. The results suggest that captivity can cause unnatural cranial development and that diet likely has a major impact on cranial morphology. Full article

A preliminary vaccination trial against the emergent pathogen, SARS-CoV-2, was completed in captive black-footed ferrets (Mustela nigripes; BFF) to assess safety, immunogenicity, and anti-viral efficacy. Vaccination and boosting of 15 BFF with purified SARS-CoV-2 S1 subunit protein produced a nearly 150-fold increase in mean antibody titers compared to pre-vaccination titers. Serum antibody responses were highest in young animals, but in all vaccinees, antibody response declined rapidly. Anti-viral activity from vaccinated and unvaccinated BFF was determined in vitro, as well as in vivo with a passive serum transfer study in mice. Transgenic mice that received BFF serum transfers and were subsequently challenged with SARS-CoV-2 had lung viral loads that negatively correlated (p < 0.05) with the BFF serum titer received. Lastly, an experimental challenge study in a small group of BFF was completed to test susceptibility to SARS-CoV-2. Despite viral replication and shedding in the upper respiratory tract for up to 7 days post-challenge, no clinical disease was observed in either vaccinated or naive animals. The lack of morbidity or mortality observed indicates SARS-CoV-2 is unlikely to affect wild BFF populations, but infected captive animals pose a potential risk, albeit low, for humans and other animals. Full article

This paper focuses on prosodic adjunction at the Prosodic Word level in a polysynthetic language. I argue that recursion at a depth of more than two levels can only be generated by a theory which requires exact correspondence between certain syntactic phrases and Prosodic Words. Such a theory is similar to Phonological Phrase correspondence in Match Theory, suggesting there is an underlying shared property between correspondence at the Prosodic Word and Phonological Phrase levels. In addition, this theory must include a constraint which prohibits recursive prosodic constituents in order to generate the attested typology of clitics across languages. The empirical focus is the prosodic structure of the verbal complex in Blackfoot (Algonquian; ISO 639-3: bla). Using phonotactic evidence I argue that the vP phase corresponds to a Prosodic Word, and that each prefix to the stem is a Prosodic Word adjunct. I then compare several theories of the syntax-prosody interface, including versions of Alignment Theory, Wrap Theory, and Match Theory. A subset of schematic candidates with one or two prefixes to a stem are used to determine which theories generate the attested typology of clitics as well as a multiply recursive Prosodic Word structure. Full article

In this paper, a complete mitochondrial genome of the critically endangered European mink Mustela lutreola L., 1761 is reported. The mitogenome was 16,504 bp in length and encoded the typical 13 protein-coding genes, two ribosomal RNA genes and 22 transfer RNA genes, and harboured a putative control region. The A+T content of the entire genome was 60.06% (A > T > C > G), and the AT-skew and GC-skew were 0.093 and −0.308, respectively. The encoding-strand identity of genes and their order were consistent with a collinear gene order characteristic for vertebrate mitogenomes. The start codons of all protein-coding genes were the typical ATN. In eight cases, they were ended by complete stop codons, while five had incomplete termination codons (TA or T). All tRNAs had a typical cloverleaf secondary structure, except tRNA^Ser(AGC) and tRNA^Lys, which lacked the DHU stem and had reduced DHU loop, respectively. Both rRNAs were capable of folding into complex secondary structures, containing unmatched base pairs. Eighty-one single nucleotide variants (substitutions and indels) were identified. Comparative interspecies analyses confirmed the close phylogenetic relationship of the European mink to the so-called ferret group, clustering the European polecat, the steppe polecat and the black-footed ferret. The obtained results are expected to provide useful molecular data, informing and supporting effective conservation measures to save M. lutreola. Full article

The black-foot disease of grapevine is one of the most destructive diseases in viticulture and it is caused by a complex of soil-borne fungi. This study aimed to identify the species associated with black-foot disease in young grapevines in vineyards of China. Fungal isolates were identified as Campylocarpon fasciculare, based on both morphological and multi-locus phylogenetic analysis of ITS, tef1–α and ß-tubulin sequence data. For the first time in China, we report Campylocarpon fasciculare associated with symptomatic young grapevines. Koch’s postulates were performed on Vitis vinifera cv. Summer Black (SB) in a greenhouse and to confirm the pathogenicity on grapevines. This work improves the knowledge of black-foot disease in Chinese vineyards and will be helpful to growers in their decisions regarding vinicultural practices, planting and disease management. Full article

Plant roots support complex microbial communities that can influence nutrition, plant growth, and health. In grapevine, little is known about the impact of abiotic stresses on the belowground microbiome. In this study, we examined the drought-induced shifts in fungal composition in the root endosphere, the rhizosphere and bulk soil by internal transcribed spacer (ITS) high-throughput amplicon sequencing (HTAS). We imposed three irrigation regimes (100%, 50%, and 25% of the field capacity) to one-year old grapevine rootstock plants cv. SO₄ when plants had developed 2–3 roots. Root endosphere, rhizosphere, and bulk soil samples were collected 6- and 12-months post-plantation. Drought significantly modified the overall fungal composition of all three compartments, with the root endosphere compartment showing the greatest divergence from well-watered control (100%). The overall response of the fungal microbiota associated with black-foot disease (Dactylonectria and “Cylindrocarpon” genera) and the potential biocontrol agent Trichoderma to drought stress was consistent across compartments, namely that their relative abundances were significantly higher at 50–100% than at 25% irrigation regime. We identified a significant enrichment in several fungal genera such as the arbuscular mycorrhizal fungus Funneliformis during drought at 25% watering regime within the roots. Our results reveal that drought stress, in addition to its well-characterized effects on plant physiology, also results in the restructuring of grapevine root microbial communities, and suggest the possibility that members of the altered grapevine microbiota might contribute to plant survival under extreme environmental conditions. Full article

(1) Background. An extensive survey of grapevine-sown cover crops and spontaneous weed flora was conducted from 2019 to 2020 in organic vineyards in six European countries (France, Italy, Romania, Slovenia, Spain, Switzerland). Our main objective was to detect and identify the presence of Cylindrocarpon-like asexual morphs species associated with black-foot disease on their roots. (2) Methods. Fungal isolations from root fragments were performed on culture media. Cylindrocarpon-like asexual morph species were identified by analyzing the DNA sequence data of the histone H3 (his3) gene region. In all, 685 plants belonging to different botanical families and genera were analyzed. Cylindrocarpon-like asexual morphs were recovered from 68 plants (9.9% of the total) and approximately 0.97% of the plated root fragments. (3) Results. Three fungal species (Dactylonectria alcacerensis, Dactylonectria torresensis, Ilyonectria robusta) were identified. Dactylonectria torresensis was the most frequent, and was isolated from many cover crop species in all six countries. A principal component analysis with the vineyard variables showed that seasonal temperatures and organic matter soil content correlated positively with Cylindrocarpon-like asexual morphs incidence. (4) Conclusions. The presence of Cylindrocarpon-like asexual morphs on roots of cover crops suggests that they can potentially act as alternative hosts for long-term survival or to increase inoculum levels in vineyard soils. Full article

There is a longstanding debate in the literature about if, and where, recursion occurs in prosodic structure. While there are clear cases of genuine recursion at the phrase level and above, there are very few convincing cases of word-level recursion. Most cases are—by definition—not recursive and instead best analyzed as different constituents (e.g., the Composite Group, Prosodic Word Group, etc.). We show that two convincing cases of prosodic word-level recursion can easily be reanalyzed without recursion if phonology and prosody are evaluated cyclically at syntactic phase boundaries. Our analysis combines phase-based spell-out and morpheme-specific subcategorization frames of Cophonologies by Phase with Tri-P Mapping prosodic structure building. We show that apparent word-level recursion is due to cyclic spell-out, and non-isomorphisms between syntactic and prosodic structure are due to morpheme-specific prosodic requirements. Full article

Aeromonas hydrophila was isolated from an African black-footed penguin (Spheniscus demersus) that died while in zoo captivity. At necropsy, the virulence of A. hydrophila appeared to be enhanced by stress, so was assessed in the presence of in vitro and in vivo stressors and serial passaging in mice. Virulence genes from the isolate were amplified by PCR. In vitro assays were conducted to test the hemolytic activity, cytotoxicity, and effect of stress hormones on A. hydrophila virulence. In vivo assays were conducted to test the stress effect on mortality of A. hydrophila-infected mice and virulence in mice. Two virulence genes coding for hemolysin (ahh1) and aerolysin (aerA) were detected, and the cytotoxic potential of the isolate was demonstrated in baby hamster kidney and Vero cells. Some or all mice inoculated with A. hydrophila and exposed to stress hormones (epinephrine and norepinephrine) or low temperature died, while mice inoculated with A. hydrophila and exposed to fasting or agitation stressors or no stressors survived. We concluded that stress can be fatal in mice experimentally infected with A. hydrophila and that serial passaging in mice dramatically enhances the virulence of A. hydrophila. Full article

The soil-borne fungus Dactylonectria torresensis is the most common causal agent of black-foot disease in Europe. However, there is a lack of understanding on how this fungus can provoke plant symptoms. In this study, we sequenced, annotated and analyzed the genomes of three isolates of D. torresensis collected from asymptomatic vine, weed and soil. Sequenced genomes were further compared to those of 27 fungal species including root and aerial pathogens, white rot degraders, indoor biodeterioration agents, saprotrophs, dark septate endophytes and mycorrhiza. Strains of D. torresensis present genomes with between 64 and 65 Mbp and with up to 18,548 predicted genes for each strain. Average Nucleotide Identity (ANI) shows that strains are different according to genome contents. Clusters of orthologous groups were compared, and clusters of genes related to necroses were particularly detected in all strains of D. torresensis (necrosis inducing peptides and proteins, and ethylene inducing peptides) as well as several genes involved in resistance against fungicides frequently used in viticulture such as copper. Interestingly, an expanded high number of genes related to carbohydrate-active enzymes were detected in each Dactylonectria strain, especially those related to glycoside hydrolases that could be involved in penetration of plant tissues or pathogenicity. An increased number of candidate genes for CAZyme classes AA9 and AA3-1 supports the ability of strains to efficiently degrade plant material. High numbers of genes of D. torresensis related to secretome and small secreted proteins were further characterized. Moreover, the presence of several gene clusters such as fujikurin-like genes was detected and were normally found in Fusariumfujikuroi, that have been linked to fungal pathogenicity. The phenotypes of the three strains investigated showed further difference in light response. We found that Dactylonectria strains have an increased number of photoreceptor encoding genes and we showed sequence alterations. Altogether, the results highlight several gene clusters present in D. torresensis strains that could be linked to endophytic lifestyle, pathogenicity, plant maceration and degradation of plant tissues as well as adaptation to soil contaminated with metals and metalloids and light response. Full article

Systematic cryo-banking of semen and testicular tissues is critical to preserve the genetic value of recently deceased or neutered black-footed ferrets (BFFs). Specifically, recovering or producing mature sperm cells from vitrified-warmed issues offers additional options in assisted reproduction. This could, in turn, enhance the genetic management of this rare and endangered species over multiple generations. The objective of the study was to evaluate structural properties, DNA fragmentation, cell viability, and germ cell composition in vitrified testicular tissues from BFFs directly after warming or after warming plus a short in vitro culture period. Tissue biopsies from five adult BFFs were either kept fresh or vitrified with a standard protocol (using dimethylsulphoxide (DMSO) and glycerol) and warmed at 50 °C for 5 s. Some of the warmed samples were then cultured in vitro for 24 h. Fresh, warmed, and warmed/cultured tissues were analyzed using different indicators: histology of seminiferous tubules, intact Sertoli cells (vimentin labeling), DNA integrity, cell viability, germ cell composition (Oct4 and Boule labeling). Percentages of intact seminiferous tubules decreased after vitrification/warming and returned to the level of fresh samples after culture. While percentages of cells labeled with vimentin, with intact DNA integrity, or proportions of viable cells were affected by vitrification/warming, they all reached similar or better levels than the fresh tissue after culture. Proportions of cells labeled with Boule antibodies also improved during in vitro culture post-warming. We demonstrated for the first time that BFF testes subjected to vitrification, rapid warming, and short in vitro culture were viable and maintained the ability to resume germ cell progression. Cryopreserved testicular tissues could potentially contribute to new strategies to enhance BFF assisted reproduction as well as conservation efforts. Full article

Chitosan is prepared by the deacetylation of chitin, the second-most abundant biopolymer in nature, and has applicability in the removal of dyes, heavy metals and radioactive waste for pollution control. In weight-reduction remedies, chitosan is used to form hydrogels with lipids and to depress the intestinal absorption of lipids. In this study, an experimental method was implemented to simulate the effect of chitosan on the adsorption of humic acid in the gastrointestinal tract. The adsorption capacity of chitosan was measured by its adsorption isotherm and analyzed using the Langmuir equation. The results showed that 3.3 grams of humic acid was absorbed by 1 gram of chitosan. The adsorption capacity of chitosan was much greater than that of chitin, diethylaminoethyl-cellulose or activated charcoal. Cellulose and carboxymethyl-cellulose, a cellulose derivative with a negative charge, could not adsorb humic acid in the gastrointestinal tract. This result suggests that chitosan entraps humic acid because of its positive charge. Full article