Effects of Contact and Ingestion Exposure to Formulated CheckMate® LBAM-F and Unformulated LBAM Mating Pheromones on Adult Worker Honeybees,Apis mellifera (Hymenoptera: Apidae)

Acute 7-day toxicity tests evaluating adverse effects from contact and ingestion exposure to light brown apple moth (LBAM) pheromones and time-released microencapsulated LBAM pheromones in CheckMate® LBAM-F (Checkmate) were conducted on newly emerged honeybees (less than 24 h old). Contact studies exposed bees to 1× and 10× the CheckMate label application rate. Ingestion studies exposed bees to CheckMate formulations, and active (pheromone) ingredients (a.i.) at 0.1%, 1.0%, and 10% concentrations by weight in solid food. Bees ingested approximately 39% of their body weight during the tests. Mortality ranged from 2–10% in three of four contact and ingestion exposure trials. Trial 1, which utilized a different feeding design, showed higher mortality in both control and test replicates (9–28%). One-way ANOVA tests indicated no significant difference in mortality between control and treatment replicates in the four trials. Bees were subjected to one-time CheckMate contact exposures of up to 0.49 mg/kg-bee, and average pheromone and formulation ingestion exposures of up to 56 (0.1%), 611 (1.0%), and 6,282 (10%) mg/kg-bee-day. LBAM pheromones and microencapsulated pheromones proved to be non-toxic to honeybees when sprayed with 10× the field application rate, or when ingested in food at concentrations of up to 10% by weight.     

The tetrameric pheromone module SteC‐MkkB‐MpkB‐SteD regulates asexual sporulation,sclerotia formation and aflatoxin production in Aspergillus flavus

For eukaryotes like fungi to regulate biological responses to environmental stimuli, various signalling cascades are utilized, like the highly conserved mitogen‐activated protein kinase (MAPK) pathways. In the model fungus Aspergillus nidulans, a MAPK pathway known as the pheromone module regulates development and the production of secondary metabolites (SMs). This pathway consists five proteins, the three kinases SteC, MkkB and MpkB, the adaptor SteD and the scaffold HamE. In this study, homologs of these five pheromone module proteins have been identified in the plant and human pathogenic fungus Aspergillus flavus. We have shown that a tetrameric complex consisting of the three kinases and the SteD adaptor is assembled in this species. It was observed that this complex assembles in the cytoplasm and that MpkB translocates into the nucleus. Deletion of steC, mkkB, mpkB or steD results in abolishment of both asexual sporulation and sclerotia production. This complex is required for the positive regulation of aflatoxin production and negative regulation of various SMs, including leporin B and cyclopiazonic acid (CPA), likely via MpkB interactions in the nucleus. These data highlight the conservation of the pheromone module in Aspergillus species, signifying the importance of this pathway in regulating fungal development and secondary metabolism.     

水椰八角铁甲不同日龄和性别成虫表皮碳氢化合物的种类及动态

水椰八角铁甲Octodonta nipae(Maulik)是一种严重为害棕榈科植物的外来入侵害虫。为了明确日龄和性别等个体因素对水椰八角铁甲成虫表皮碳氢化合物的影响,本研究采用化学萃取法和气相色谱-质谱联用技术,分析了不同日龄的水椰八角铁甲雌、雄成虫表皮碳氢化合物的组成和含量。结果表明,水椰八角铁甲成虫的表皮碳氢化合物由21种C_(18)-C_(30)的碳氢化合物构成,包括11种正构烷烃、2种单甲基烷烃、1种多甲基烷烃、3种正构烯烃、1种单甲基烯烃和3种环烷烃。不同日龄和性别成虫的表皮碳氢化合物的组成种类相同,但是一些组分的含量有明显变化,例如1 d、5 d、10 d和15 d雌、雄成虫之间正二十四烷、正二十六烷、2-甲基十九烷和1-二十烯的含量存在显著差异。水椰八角铁甲成虫表皮碳氢化合物具有性二型,如性成熟雄虫的表皮的正二十烷、正二十一烷、正二十四烷、正二十六烷、2-甲基十九烷、10-甲基二十烷、1-二十烯和1-二十二烯的含量显著高于性成熟雌虫。本文的研究结果可为检验检疫过程中水椰八角铁甲近缘种,以及日龄、性别和性成熟的快速鉴定提供支持。     

Short communication: administering an appeasing substance to Bos indicus-influenced beef cattle at weaning and feedlot entry

The bovine appeasing substance (BAS) is expected to have calming effects in cattle experiencing stressful situations. Therefore, this study investigated the impacts of BAS administration during two of the most stressful events within beef production systems: weaning and feedlot entry. In experiment 1, 186 Bos indicus-influenced calves (73 heifers, 113 bulls) were weaned at 211 ± 1 days of age (day 0). At weaning, calves were ranked by sex and BW, and assigned to receive BAS (Nutricorp, Araras, SP, Brazil; n = 94) or water (CON; n = 92). Treatments (5 ml) were topically applied to the nuchal skin area of each animal. Calf BW was recorded and samples of blood and tail-switch hair were collected on days 0, 15 and 45. Calves that received BAS had greater (P < 0.01) BW gain from day 0 to 15 compared with CON. Overall BW gain (days 0 to 45) and BW on days 15 and 45 were also greater (P ≤ 0.03) in BAS v. CON. Plasma haptoglobin concentration was less (P < 0.01) in BAS v. CON on day 15, whereas cortisol concentrations in plasma and tail-switch hair did not differ between treatments (P ≥ 0.13). In experiment 2, 140 B. indicus-influenced bulls (∼27 months of age) from 2 different pasture-based systems (70 bulls/origin) were transported to a commercial feedlot (≤ 200-km transport; day -1). On day 0, bulls were ranked by source and BW, and assigned to receive BAS (n = 70) or CON (n = 70) and the same sampling procedures as in experiment 1. Bulls receiving BAS had greater (P = 0.04) BW gain from day 0 to 15, but less (P < 0.01) BW gain from day 15 to 45 compared to CON. No other treatment effects were detected (P > 0.14). Therefore, BAS administration to beef calves alleviated the haptoglobin response associated with weaning, and improved calf growth during the subsequent 45 days. Administration of BAS to beef bulls at feedlot entry improved BW gain during the initial 15 days, but these benefits were not sustained throughout the 45-day experiment.     

Biotic and abiotic factors driving the diversification dynamics of Crocodylia

Species diversity patterns are governed by complex interactions among biotic and abiotic factors over time and space, but are essentially the result of the diversification dynamics (differential speciation and extinction rates) over the long-term evolutionary history of a clade. Previous studies have suggested that temporal variation in global temperature drove long-term diversity changes in Crocodylia, a monophyletic group of large ectothermic organisms. We use a large database of crocodylian fossil occurrences (192 spp.) and body mass estimations, under a taxic approach, to characterize the global diversification dynamics of crocodylians since the Cretaceous, and their correlation with multiple biotic and abiotic factors in a Bayesian framework. The diversification dynamic of crocodylians, which appears to have originated in the Turonian (c. 92.5 Ma), is characterized by several phases with high extinction and speciation rates within a predominantly low long-term mean rate. Our results reveal long-term diversification dynamics of Crocodylia to be a highly complex process driven by a combination of biotic and abiotic factors which influenced the speciation and extinction rates in dissimilar ways. Higher crocodylian extinction rates are related to low body mass disparity, indicating selective extinctions of taxa at both ends of the body mass spectrum. Speciation rate slowdowns are noted when the diversity of the clade is high and the warm temperate climatic belt is reduced. Our finding supports the idea that temporal variations of body mass disparity, self-diversity, and the warm climate belt size provided more direct mechanistic explanations for crocodylian diversification than do proxies of global temperature.     

Genome-Wide Association Analysis Identifies a Genetic Basis of Infectivity in a Model Bacterial Pathogen

Knowledge of the genetic architecture of pathogen infectivity and host resistance is essential for a mechanistic understanding of coevolutionary processes, yet the genetic basis of these interacting traits remains unknown for most host–pathogen systems. We used a comparative genomic approach to explore the genetic basis of infectivity in Pasteuria ramosa, a Gram-positive bacterial pathogen of planktonic crustaceans that has been established as a model for studies of Red Queen host–pathogen coevolution. We sequenced the genomes of a geographically, phenotypically, and genetically diverse collection of P. ramosa strains and performed a genome-wide association study to identify genetic correlates of infection phenotype. We found multiple polymorphisms within a single gene, Pcl7, that correlate perfectly with one common and widespread infection phenotype. We then confirmed this perfect association via Sanger sequencing in a large and diverse sample set of P. ramosa clones. Pcl7 codes for a collagen-like protein, a class of adhesion proteins known or suspected to be involved in the infection mechanisms of a number of important bacterial pathogens. Consistent with expectations under Red Queen coevolution, sequence variation of Pcl7 shows evidence of balancing selection, including extraordinarily high diversity and absence of geographic structure. Based on structural homology with a collagen-like protein of Bacillus anthracis, we propose a hypothesis for the structure of Pcl7 and the physical location of the phenotype-associated polymorphisms. Our results offer strong evidence for a gene governing infectivity and provide a molecular basis for further study of Red Queen dynamics in this model host–pathogen system.     

High yield expression of biologically active recombinant full length human tuftelin protein in baculovirus-infected insect cells

Tuftelin is an acidic protein expressed at very early stages of mouse odontogenesis. It was suggested to play a role during epithelial–mesenchymal interactions, and later, when enamel formation commences, to be involved in enamel mineralization. Tuftelin was also detected in several normal soft tissues of different origins and some of their corresponding cancerous tissues. Tuftelin is expressed in low quantities, and undergoes degradation in the enamel extracellular matrix. To investigate the structure and function of tuftelin, the full length recombinant human tuftelin protein was produced. The full length human tuftelin cDNA was cloned using Gateway^TM recombination into the Bac-to-Bac^TM system compatible transfer vector pDest10. This vector adds a hexahistidine tag to the N-terminus of the expressed protein, enabling one-step affinity purification on nickel column. The recombinant human tuftelin protein was transposed into the bacmid and expressed in Spodoptera frugiperda (Sf9) insect cells. The yield of the purified, his-tagged recombinant full length human Tuftelin (rHTuft⁺) was 5–8 mg/L culture. rHTuft⁺ was characterized by SDS–PAGE, Western blot, ESI-TOF spectrometry, restriction mapping and MS/MS sequencing. The availability of the purified, full length recombinant human tuftelin protein opened up the possibility to investigate novel functions of tuftelin. Application of rHTuft⁺ agarose beads onto embryonic mouse mandibular explants caused changes in the surrounding epithelial cells, including morphology, orientation and spatial organization. Further studies using DiI labeling, revealed that rHTuft⁺, placed on the tooth germ region, brought about recruitment of adjacent embryonic mesenchymal cells. These findings support the hypothesis that tuftelin plays an important role during embryogenesis.     

Abdominal odours of young,low-ranking European rabbit mothers are less attractive to pups: an experiment with animals living under natural breeding conditions

Lactating rabbit females (Oryctolagus cuniculus) emit abdominal, ventral odour cues that are involved in guiding the pups to the nipples. We tested the impact of the females’ social rank and age on the emission of this odour signal; both factors are highly collinear in wild rabbits and we did not aim to disentangle them in our study. We predicted that the abdominal odours of young, low-ranking females, which are usually more stressed and have a comparatively lower body condition, should be less attractive to pups than those of older, high-ranking females. For the experiment, we caught females from a population of wild-type European rabbits living in a large field enclosure. Pairs of adult females were placed in boxes with wire-mesh floor fixed over an arena with three rabbit pups aged 4–7 days old. First, we evaluated the effectiveness of our two-choice design by testing lactating females (LF) against non-lactating (NLF) controls (n = 17 pairs). In accordance with recent studies, the pups showed a clear preference for lactating females by means of a higher exploration time and more local searching under the ventral region of these females. In a second experiment, the pups chose between lactating 1-year-old, low-ranking females (Y/LRF) and older, high-ranking females (O/HRF) (n = 10 pairs). These latter assays suggested a comparatively lower attractiveness of Y/LRF females to pups searching for milk. Although the pups showed no significant preference in exploration behaviour, they preferably directed local searching events to the abdominal region of O/HRF females. In conclusion, our study suggests that mother–offspring chemosensory communication is modified by the mother’s age and social rank, i.e., by a complex of factors that are strongly linked in rabbit females in the wild.