Grain,pellet and wax block bait take by the house mouse (Mus musculus) and non‐target species: implications for mouse eradications on coral cay islands

Introduced rodents have been eradicated from large numbers of offshore islands using toxic baits; however, toxic baits have been linked with negative impacts on non‐target species. The present study assessed the bait take of target (house mouse, Mus musculus) and non‐target (buff banded rail, Rallus philippensis) animals on Northwest and Heron Islands in the Great Barrier Reef. Three non‐toxic bait formulations (wax block, pellet and grain) were tested and each was applied at 1 kg ha^?1 in six treatment grids. The tracks of animals visiting the baits were identified using 30 tracking stations per treatment grid. A tracking station consisted of a track‐board placed in the centre of a sand‐pad. Mean bait take differed significantly between the formulations: birds took more grain bait than wax block bait; mice took more wax block than grain bait. Both mice and birds were equally selective of pellet bait. Thus, the findings indicate that wax blocks are the most suitable formulation for future baiting programs to eradicate mice on these and other islands.     

Efficacy of anticoagulant-free alternative bait products against house mice (Mus musculus) and brown rats (Rattus norvegicus)

The attractiveness and efficacy of four anonymous anticoagulant-free alternative rodenticides (active ingredients cellulose or plaster) were tested on wild strain groups of house mice (Mus musculus Linnaeus) and brown rats (Rattus norvegicus Berkenhout) in laboratory choice and no-choice experiments. In no-choice tests, the rodenticide product was offered ad libitum to the animals for 21 days (mice) and 10 days (rats), respectively. In choice tests, the rodenticide product and non-poisoned food were offered for 28 days (mice) and 14 days (rats). Two products with cellulose (C1 and C2) as the active ingredient were tested against both mice and rats. In two no-choice tests with mice, all animals died within 14 days (C1) and 21 days (C2); in a third no-choice test, 11% (2/18) of mice survived (C1: 21 days). In all three tests, numerous incidents of cannibalism were observed. The attractiveness of cellulose baits was tested for one product (C1) in three trials against normal food pellets. In all trials, bait consumption was low and all mice survived (15–34 days). Two no-choice trials with cellulose baits (C1) were conducted on groups of Rattus norvegicus; 11 of 12 rats survived the bait feeding period and no cannibalism or sign of aggression between conspecifics was observed. Two plaster-based (CaSO₄·0.5H₂O) products (P1 and P2) were tested against both mice and rats; in no-choice tests, only one mouse (n= 20) and no rats (n= 8) were killed, although the baits proved to be attractive for rats and mice in choice tests. The results of these experiments suggest that neither cellulose-based nor plaster-based rodenticides are suitable agents for the control of Mus musculus and Rattus norvegicus.     

兔原始生殖细胞体外培养的研究

旨在探索兔原始生殖细胞（primordial germ cells,PGCs）体外分离培养的最佳条件,从而建立成熟稳定的兔PGCs体外分离培养方法。本研究首先通过两种不同的体外分离法（胰酶消化法、机械法）和3种不同的传代法（胰酶消化法、机械法、连同饲养层消化法）探索第14~18天胎兔的PGCs体外分离传代的最佳方式,另将培养液分为A、B、C、D 4组,以D组为对照组,探究不同细胞因子浓度对兔PGCs形态变化及集落形成的影响。其次,利用碱性磷酸酶染色（alkaline phosphatase staining,AKP）法对兔PGCs进行鉴定染色;实时荧光定量（real-time polymerase chain reaction,RT-PCR）检测转录因子Oct-4的表达。结果表明,机械法分离得到的兔PGCs集落数量是酶消化法分离的2.2倍,而兔PGCs经不同的消化法传代发现,酶消化法、连同饲养层消化法、机械法在兔胚胎成纤维细胞（rabbit embryo fibroblast,REF）饲养层上分别成功传至P2、P2、P4代。B组PGCs培养液（基础液+10%胎牛血清（fetal bovine serum,FBS）+2 ng·mL^-1转化生长因子-β1（transforming growth factor-β1,TGF-β1）+4 ng·mL^-1碱性成纤维细胞生长因子（basic fibroblast growth factor, bFGF）+10 ng·mL^-1白血病抑制因子（leukemia inhibitory factor,LIF））所获得的集落数量最多,具有较好的集落形态,保持未分化的时间较长。AKP染色结果显示,PGCs集落呈红黑色; RT-PCR结果显示,体外分离培养的兔PGCs表达转录因子Oct-4。结果显示,兔原代PGCs最适采用机械分离法和机械传代法进行体外分离传代,适宜浓度的细胞因子添加至兔PGCs培养液中有利于兔PGCs在体外保持较多的集落数量和较长时间的未分化状态。本研究通过筛选和优化兔PGCs体外培养方法,为进一步建立稳定成熟兔PGCs细胞系奠定技术基础。     

家兔矮小同源盒基因(Shox2)的克隆和表达分析

为了研究新西兰白兔矮小同源盒基因(Shox2)序列,预测其结构和功能,以及研究Shox2基因的时空表达特征,本研究克隆了新西兰白兔Shox2基因全长CDS区,分析其编码蛋白的同源性、亲疏水性、二级和三级结构特点、系统进化树等生物信息学特征,并用Western blot法验证融合蛋白TrxA-Shox2。分别在胚胎中期(E20.5d)、胚胎后期(E28.5d)、幼龄期(出生后10d)、成年期(A)4个时间点采集3只动物的肾、大血管、真皮、心、大脑、肝、肺、脾、肌肉、胆囊共10种组织,采用实时荧光定量PCR技术研究Shox2基因的时空表达图谱。结果,成功克隆了新西兰白兔Shox2基因(登录号:KP726285)。Shox2基因CDS区全长666bp,翻译221个氨基酸。生物信息学分析显示,Shox2为碱性、不稳定蛋白,Shox2蛋白氨基酸二级结构中α-螺旋区域占61.99%,无规则卷曲区域占33.48%,延伸链占4.52%。Western blot验证了融合蛋白TrxA-Shox2的表达。家兔Shox2蛋白氨基酸序列与眼镜猴、猩猩、褐家鼠、人的氨基酸序列同源性较高,说明Shox2蛋白在进化中较保守。Shox2基因mRNA在很多器官都有表达,在E20.5d的家兔胚胎中,Shox2mRNA在肾、肌肉和胆囊的表达量高,在E28.5d时表达量较高的是脾、心,幼龄期时表达量较高的是真皮、肝,成年期时表达量较高的是真皮、脾。本研究成功克隆了新西兰白兔Shox2基因,预测了其结构和功能。Shox2基因mRNA在很多器官都有表达,不同的组织和不同时间特征的表达量变化可能与Shox2功能高度相关,并受到严格的调控。     

Evolution of rabbit haemorrhagic disease virus (RHDV) in the European rabbit (Oryctolagus cuniculus) from the Iberian Peninsula

To date information on rabbit haemorrhagic disease virus (RHDV) in Spain and Portugal has been scarce, although the disease is endemic and continues to have a considerable impact on species conservation and hunting industry. We analysed RHDVs obtained between 1994 and 2007 at different geographic locations in Portugal (40 samples), Spain (3 samples) and France (4 samples) from wild European rabbits (Oryctolagus cuniculus) that succumbed to the disease. Phylogenetic analyses based on partial VP60 gene sequences allowed a grouping of these RHDVs into three groups, termed "Iberian" Groups IB1, IB2 and IB3. Interestingly, these three Iberian groups clustered separately, though not far from earlier RHDVs of Genogroup 1 (containing e.g., strain "AST89"), but clearly distinct from globally described RHDV strains of Genogroups 2-6. This result, supported by a bootstrap value of 76%, gives rise to the hypothesis that the virus evolved independently since its introduction to wild rabbit populations on the Iberian Peninsula, with the Pyrenees acting as a natural barrier to rabbit and hence to virus dispersal. No differences were observed in RHDV sequences obtained from geographic regions where the rabbit subspecies O. c. algirus prevails compared with those obtained from O. c. cuniculus.     

Dissection of white matter association fasciculi in bovine (Bos taurus), pig (Sus scrofa domesticus) and rabbit (Oryctolagus cuniculus) brains

The cerebral fasciculi (association, commissural and projection) pass through the cerebral white matter in organized groups connecting regions, hemispheres, gyri, areas and brain lobes to each other. The study can be done in vivo through diffusion tensor imaging (DTI) but presenting some technical problems. The post-mortem study by dissection allows to have a clearer view of its location, path and connections. In this work, we dissect, identify and compare the fasciculi of association of the white matter of the dorsolateral face of bovine hemispheres (Bos taurus), pig hemispheres (Sus scrofa domesticus) and rabbit hemispheres (Oryctolagus cuniculus), applying the Klingler´s technique. In 30 cerebral hemispheres (10 of each species, five right and five left), we applied the Klingler technique to identify and isolate the occipitofrontal fasciculus, uncinate fasciculus, inferior longitudinal fasciculus and superior longitudinal fasciculus; we established its location by fixing landmarks, and determined the difference in its length and width between the right and left hemispheres as well as between species using the statistical tests of t-student and one-way ANOVA. We identify the gyri, sulci and fasciculi of the dorsolateral surface of the cerebral hemispheres of the three species. We found statistically significant differences in several fasciculi between species principally in the occipitofrontal fasciculus. The preparation of the brains through the modified Klingler technique allowed a successful identification of the fascicules of association of the dorsolateral face of the cerebral hemispheres and the empowerment of these animal models for future research work in this field.     

Morphological changes of physeal cartilage and secondary ossification centres in the developing femur of the house mouse (Mus musculus): A micro‐CT based study

In mammals, long bones are formed by ossification of a cartilaginous mould during early stages of development, through the formation of structures called the primary ossification centre, the secondary ossification centres (SOCs) and the physeal cartilages (PCs). The PC is responsible for long bone growth. The morphology of the PC and the SOCs varies during different stages of femoral growth. In this respect, several details involving the process of murine femoral development are lacking. In the present study, a morphological characterization of femur development from the embryonic period to adulthood in mice was studied using micro‐computed tomography (micro‐CT). To achieve this aim, femora were collected at embryonic day (E) 14.5, E16.5 and E18.5 and at postnatal day (P)1, P7, P14, P35, P46 and P52. CT images were obtained using a micro‐CT scanner (X‐SkyScan 1172; Micro Photonics) and analysed using the micro‐CT 3D visualization software Mimics (Materialise NV, Leuven, Belgium) and NRecon (Micro Photonics). The results of the present study revealed that the femur and its PCs and SOCs undergo morphological changes during different stages of development, including changes in their shape as well as position and thickness. These changes may be due to the response of the femur to mechanical loads imposed by muscle surrounding the bone during these stages of development. The result of the present study is important to improve our knowledge related to ossification and growth patterns of mouse femur during development.     

Population increase and changes in behavior and morphology in the Critically Endangered Redonda ground lizard (Pholidoscelis atratus) following the successful removal of alien rats and goats

Redonda is a small volcanic Caribbean island that is home to at least 4 endemic lizard species, including the Critically Endangered ground lizard (Pholidoscelis atratus). Black rats (Rattus rattus) and domestic goats (Capra hircus) were introduced to the island at some time after its discovery by Europeans in the late 1500s. They had a devastating effect on the island, resulting in the loss of nearly all trees and most of the ground vegetation. Point count surveys of P. atratus in 2012 indicated low densities, and the invasive rats were observed hunting and preying on the lizards. Both populations of rats and goats were successfully removed in 2017 as part of an ecological restoration program, and native vegetation and invertebrate populations have increased rapidly since. Population surveys in 2017, 2018, and 2019 show the lizard population has increased by more than sixfold. In 2017, as rats and goats were being removed, we evaluated the morphology and escape behavior of this species and repeated these measurements 1 year later. We observed that P. atratus had become bolder, with a reduced flight distance. We also detected changes in limb morphology related to locomotion and suggest possible explanations that will need to be further investigated in the future. These results show how the removal of invasive species can rapidly affect lizard population recovery and behavior, potentially restoring island ecosystems to their pre-human interference dynamics.