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The shells of large benthic foraminifers (LBF) are key contributors to the development and maintenance of coastal landforms in the Pacific as well as to Paleogene and Miocene carbonates deposited along the Neotethys Seaway and tropical Pacific islands. The current study assessed growth and fecundity of two species, Marginopora vertebralis and Amphistegina lobifera, collected from sites in Viti Levu, Fiji, based on shell diameter, shell weight, fecundity, and survival. Specimens were cultured without supplemental nutrients or food for 15 months under controlled laboratory conditions. Physicochemical parameters, including salinity, alkalinity, pH, and temperature, varied by

Psoroptes are a non-burrowing, ectoparasitic, mange-causing mite that has been documented in American bighorn sheep populations throughout the 19th and 20th centuries; however, it was not seen on Canadian bighorn sheep until 2006. The aim of this study was to determine the potential source of the Psoroptes outbreak in Canadian bighorn sheep. Morphological and molecular analyses were used to compare mites recovered from outbreak-associated bighorn sheep, pet rabbits in Canada, and on historically infested bighorn sheep in the USA. The results revealed that Psoroptes acquired from the Canadian and outbreak-associated American bighorn sheep were morphologically more similar to those collected from rabbits than mites on historically infested bighorn sheep. Outer opisthosomal setae lengths measured an average of 81.7 μm (±7.7 μm) in outbreak associated bighorn mites, 88.9 μm (±12.0 μm) in rabbit mites and 151.2 μm (±16.6 μm) in historically infested bighorn mites. The opisthosomal lobe morphology of bighorn mites in the outbreak herds was also more similar to that of rabbit mites, previously described as P. cuniculi, than historically infested bighorn mites, which match previous descriptions of P. ovis. This finding was supported by DNA sequence data of the mitochondrial cytochrome B gene. This is the first report of Psoroptes of the rabbit ecotype on bighorn sheep. The morphological and molecular data therefore support the hypothesis that the source of Psoroptes outbreak in Canadian bighorn sheep represented a disease spillover event from rabbits rather than transmission from infested American bighorn sheep populations.

Many copy number variants (CNVs) confer risk for the same range of neurodevelopmental symptoms and psychiatric conditions including autism and schizophrenia. Yet, to date neuroimaging studies have typically been carried out one mutation at a time, showing that CNVs have large effects on brain anatomy. Here, we aimed to characterize and quantify the distinct brain morphometry effects and latent dimensions across 8 neuropsychiatric CNVs. We analyzed T1-weighted MRI data from clinically and non-clinically ascertained CNV carriers (deletion/duplication) at the 1q21.1 (n = 39/28), 16p11.2 (n = 87/78), 22q11.2 (n = 75/30), and 15q11.2 (n = 72/76) loci as well as 1296 non-carriers (controls). Case-control contrasts of all examined genomic loci demonstrated effects on brain anatomy, with deletions and duplications showing mirror effects at the global and regional levels. Although CNVs mainly showed distinct brain patterns, principal component analysis (PCA) loaded subsets of CNVs on two latent brain dimensions, which explained 32 and 29% of the variance of the 8 Cohen’s d maps. The cingulate gyrus, insula, supplementary motor cortex, and cerebellum were identified by PCA and multi-view pattern learning as top regions contributing to latent dimension shared across subsets of CNVs. The large proportion of distinct CNV effects on brain morphology may explain the small neuroimaging effect sizes reported in polygenic psychiatric conditions. Nevertheless, latent gene brain morphology dimensions will help subgroup the rapidly expanding landscape of neuropsychiatric variants and dissect the heterogeneity of idiopathic conditions.

Leptospermum scoparium (‘Mānuka’) is native to New Zealand and South Eastern Australia. Throughout this range the species typically exists as a diploid. Only a single tetraploid accession h...

Membraneless organelles containing the enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) are a common feature of organisms utilizing CO2 concentrating mechanisms to enhance photosynthetic carbon acquisition. In cyanobacteria and proteobacteria, the Rubisco condensate is encapsulated in a proteinaceous shell, collectively termed a carboxysome, while some algae and hornworts have evolved Rubisco condensates known as pyrenoids. In both cases, CO2 fixation is enhanced compared with the free enzyme. Previous mathematical models have attributed the improved function of carboxysomes to the generation of elevated CO2 within the organelle via a colocalized carbonic anhydrase (CA) and inwardly diffusing HCO3−, which have accumulated in the cytoplasm via dedicated transporters. Here, we present a concept in which we consider the net of two protons produced in every Rubisco carboxylase reaction. We evaluate this in a reaction–diffusion compartment model to investigate functional advantages these protons may provide Rubisco condensates and carboxysomes, prior to the evolution of HCO3− accumulation. Our model highlights that diffusional resistance to reaction species within a condensate allows Rubisco-derived protons to drive the conversion of HCO3− to CO2 via colocalized CA, enhancing both condensate [CO2] and Rubisco rate. Protonation of Rubisco substrate (RuBP) and product (phosphoglycerate) plays an important role in modulating internal pH and CO2 generation. Application of the model to putative evolutionary ancestors, prior to contemporary cellular HCO3− accumulation, revealed photosynthetic enhancements along a logical sequence of advancements, via Rubisco condensation, to fully formed carboxysomes. Our model suggests that evolution of Rubisco condensation could be favored under low CO2 and low light environments.

Cyclic electron flow (CEF) around photosystem I (PSI) is essential for generating additional ATP and enhancing efficient photosynthesis. Accurate estimation of CEF requires knowledge of the fractions of absorbed light by PSI (fI) and PSII (fII), which are only known for a few model species such as spinach. No measures of fI are available for C4 grasses under different irradiances. We developed a new method to estimate (1) fII in vivo by concurrently measuring linear electron flux through both photosystems $$\left( {{\text{LEF}}_{{{\text{O}}_{ 2} }} } \right)$$LEFO2 in leaf using membrane inlet mass spectrometry (MIMS) and total electron flux through PSII (ETR2) using chlorophyll fluorescence by a Dual-PAM at low light and (2) CEF as ETR1—$${\text{LEF}}_{{{\text{O}}_{ 2} }}$$LEFO2. For a C3 grass, fI was 0.5 and 0.4 under control (high light) and shade conditions, respectively. C4 species belonging to NADP-ME and NAD-ME subtypes had fI of 0.6 and PCK subtype had 0.5 under control. All shade-grown C4 species had fI of 0.6 except for NADP-ME grass which had 0.7. It was also observed that fI ranged between 0.3 and 0.5 for gymnosperm, liverwort and fern species. CEF increased with irradiance and was induced at lower irradiances in C4 grasses and fern relative to other species. CEF was greater in shade-grown plants relative to control plants except for C4 NADP-ME species. Our study reveals a range of CEF and fI values in different plant functional groups. This variation must be taken into account for improved photosynthetic calculations and modelling.

A prior multilocus sequence typing (MLST) study reported that Mycoplasma bovis isolates from North American bison possess sequence types (STs) different from those found among cattle. The 42 bison isolates evaluated were obtained in 2007 or later, whereas only 19 of 94 (~20%) of the available cattle isolates, with only 1 from North America, were from that same time. We compared STs of additional, contemporary, North American cattle isolates with those from bison, as well as isolates from 2 North American deer, all originating during the same timeframe, to more definitively assess potential strain-related host specificity and expand our understanding of the genetic diversity of M. bovis. From 307 isolates obtained between 2007 and 2017 (209 from cattle, 96 from bison, 2 from deer), we identified 49 STs, with 39 found exclusively in cattle and 5 exclusively in bison. Four STs were shared between bison and cattle isolates; one ST was found in cattle and in a deer. There was no clear association between ST and the health status of the animal of origin. An MLST-based phylogeny including 41 novel STs identified in our study reveals that STs found in bison fall within several divergent lineages that include STs found exclusively in cattle.

Purpose Published literature on growth, voluntary intake, and nutrient digestion and metabolism of North American bison (Bison bison) was reviewed to create an information base for implementing management strategies. Sources The information came from refereed scientific journals, published textbooks, experiment station reports, and PhD dissertations. Synthesis After narrowly avoiding extinction in the 1880s, about 400,000 North American bison are in the United States and Canada today. Voluntary intake and growth are depressed during the winter and rebound during the following spring and summer. Domestic cattle and bison share similar grazing behavior, intake capacity, and ruminal fermentation characteristics. Greater DM digestion coefficients for bison compared with cattle appear to result from less DMI for bison than cattle in the studies. With the exceptions of red blood cell quantity and characteristics, and greater glucose and BUN concentrations, bison and cattle have similar blood histology profiles, enzyme activities, and mineral and metabolite concentrations. Conclusions and Applications Available forage biomass and nutritive value, not capacity to eat, will limit acquisition of needed nutrition by grazing bison. Management of domestic herds should take into consideration the annual “winter slump” in intake and growth. Any putative advantage in fiber fermentation capacity in bison compared with cattle does not appear to be due to differences in ruminal microbial populations.In feedlots, bison consumed feed DM equal to about 2.5% of BW, gained 750 g/d, and converted feed to gain at 9% efficiency.

Coral resilience to ocean acidification is largely determined by the degree of physiological control corals can exert on their calcifying fluid carbonate chemistry. In this study, the boron isotopic composition (δ11B) of a Porites colony growing on a reef flat on Kiritimati Island in the equatorial central Pacific is examined to quantify the sensitivity of calcifying fluid pH (pHcf) to ambient environmental conditions. Skeletal δ11B along the growth axis of one annual growth band was determined with bulk analysis and by laser ablation (LA) MC‐ICP‐MS. Furthermore, the oxygen and carbon isotopic composition, trace element ratios, and skeletal density were quantified. Sclerochronological data were interpreted in the context of simultaneous recordings of reef flat seawater pH (pHsw), temperature, salinity, and water depth, and by measurements of these parameters on the fore‐reef. A recent model of pHcf upregulation, after optimization with seasonally resolved data, was used to simulate pHcf variability on a diurnal scale. Results showed that on a seasonal scale, Porites pHcf is upregulated compared to ambient seawater: both bulk and LA‐MC‐ICP‐MS derived δ11B resulted in a mean pHcf of 8.35 pH units. Calcifying fluid pH upregulation primarily followed variations in seawater temperatures, that is likely related to the control of temperature on calcification rate. On the reef flat, the diurnal range in pHsw was substantially higher (0.29 pH units) than on the fore‐reef (0.07 pH units). However, model results suggest that the high diurnal variability in reef flat pHsw resulted only in a limited variability in Porites pHcf.

Background Mycoplasma bovis causes mastitis, otitis, pneumonia and arthritis in cattle and is a major contributor to bovine respiratory disease complex. Around the year 2000, it emerged as a significant threat to the health of North American bison. Whether healthy bison are carriers of M. bovis and when they were first exposed is not known. To investigate these questions we used a commercially available ELISA that detects antibodies to M. bovis to test 3295 sera collected from 1984 through 2019 from bison in the United States and Canada. Results We identified moderately to strongly seropositive bison from as long ago as the late 1980s. Average seroprevalence over the past 36 years is similar in the United States and Canada, but country-specific differences are evident when data are sorted by the era of collection. Seroprevalence in the United States during the pre-disease era (1999 and prior) was significantly higher than in Canada, but was significantly lower than in Canada during the years 2000–2019. Considering individual countries, seroprevalence in the United States since the year 2000 dropped significantly as compared to the years 1985–1999. In Canada the trend is reversed, with seroprevalence increasing significantly since the year 2000. ELISA scores for sera collected from free-ranging bison do not differ significantly from scores for sera from more intensively managed animals, regardless of the era in which they were collected. However, seroprevalence among intensively raised Canadian bison has nearly doubled since the year 2000 and average ELISA scores rose significantly. Conclusions Our data provide the first evidence that North American bison were exposed to M. bovis many years prior to the emergence of M. bovis-related disease. Patterns of exposure inferred from these results differ in the United States and Canada, depending on the era under consideration. Our data further suggest that M. bovis may colonize healthy bison at a level sufficient to trigger antibody responses but without causing overt disease. These findings provide novel insights as to the history of M. bovis in bison and will be of value in formulating strategies to minimize the impact of mycoplasmosis on bison health and production.

Dinoflagellates exist in symbiosis with a number of marine invertebrates including giant clams, which are the largest of these symbiotic organisms. The dinoflagellates (Symbiodinium sp.) live intercellularly within tubules in the mantle of the host clam. The transport of inorganic carbon (Ci) from seawater to Symbiodinium (=zooxanthellae) is an essential function of hosts that derive the majority of their respiratory energy from the photosynthate exported by the zooxanthellae. Immunolocalisation studies show that the host has adapted its physiology to acquire, rather than remove CO2, from the haemolymph and clam tissues. Two carbonic anhydrase (CA) isoforms (32 and 70 kDa) play an essential part in this process. These have been localised to the mantle and gill tissues where they catalyse the interconversion of HCO3- to CO2, which then diffuses into the host tissues. The zooxanthellae exhibit a number of strategies to maximise Ci acquisition and utilisation. This is necessary as they express a form II Rubisco that has poor discrimination between CO2 and O2. Evidence is presented for a carbon concentrating mechanism (CCM) to overcome this disadvantage. The CCM incorporates the presence of a light-activated CA activity, a capacity to take up both HCO3-and CO2, an ability to accumulate an elevated concentration of Ci within the algal cell, and localisation of Rubisco to the pyrenoid. These algae also express both external and intracellular CAs, with the intracellular isoforms being localised to the thylakoid lumen and pyrenoid. These results have been incorporated into a model that explains the transport of Ci from seawater through the clam to the zooxanthellae.

Hornworts (Anthocerotophyta) are the only group of land plants with pyrenoid-containing chloroplasts. CO2 exchange and carbon isotope discrimination values (Δ13C) values have previously demonstrated the presence of a CO2 concentrating mechanism (CCM) in some pyrenoid-containing species. We have examined hornwort CCM function by using a combined fluorometer/mass spectrometer based technique to compare pyrenoid-containing (PhaeocerosProsk. and Notothylas Sull.) and pyrenoid-lacking (Megaceros Campbell) hornworts, with the liverwort Marchantia polymorphaL. that has standard C3 photosynthesis and a thalloid growth form similar to hornworts. We found that Notothylas has more CCM activity than Phaeoceros, and that Megaceros has the least CCM activity. Notothylas and Phaeoceros had compensation points from 11–13 parts per million (ppm) CO2, lower K0.5(CO2) than Marchantia, negligible photorespiration, and they accumulate a pool of dissolved inorganic carbon (DIC) between 19–108 nmol mg–1 chlorophyll. Megaceroshad an intermediate compensation point of 31 ppm CO2 (compared with 64 ppm CO2 in Marchantia), a lower K0.5(CO2) than Marchantia, and some photorespiration, but no DIC pool. We also determined the catalytic rate of carboxylation per active site of Rubisco for all four species (Marchantia, 2.6 s–1; Megaceros, 3.3 s–1; Phaeoceros, 4.2 s–1; Notothylas 4.3 s-1), and found that Rubisco content was 3% of soluble protein for pyrenoid-containing species, 4% for Megaceros and 8% for Marchantia.

Cyanobacteria have developed an effective photosynthetic CO2 concentrating mechanism (CCM) for improving the efficiency of carboxylation by a relatively inefficient Rubisco. The development of this CCM was presumably in response to the decline in atmospheric CO2 levels and rising O2, both of which were triggered by the development of oxygenic photosynthesis by cyanobacteria themselves. In the past few years there has been a rapid expansion in our understanding of the mechanism and genes responsible for the CCM. In addition, there has been a recent expansion in the availability of complete cyanobacterial genomes, thus increasing our potential to examine questions regarding both the evolution and diversity of components of the CCM across cyanobacteria. This paper considers various CCM and photosynthesis gene components across eight cyanobacteria where significant genomic information is available. Significant conclusions from our analysis of the distribution of various genes indicated the following. Firstly, cyanobacteria have developed with two types of carboxysomes, and this is correlated with the form of Rubisco present. We have coined the terms α-cyanobacteria to refer to cyanobacteria containing Form 1A Rubisco and α-carboxysomes, and β-cyanobacteria having Form 1B Rubisco and β-carboxysomes. Secondly, there are two NDH-1 CO2 uptake systems distributed variably, withProchlorococcus marinus species appearing to lack this CO2 uptake system. There are at least two HCO3– transport systems distributed variably, with some α-cyanobacteria having an absence of systems identified in β-cyanobacteria. Finally, there are multiple forms of carbonic anhydrases (CAs), but with only β-carboxysomal CA having a clearly shown role at present. The α-cyanobacteria appear to lack a clearly identifiable carboxysomal CA. A pathway for the evolution of cyanobacterial CCMs is proposed. The acquisition of carboxysomes triggered by the rapid decline of atmospheric CO2 in the Phanerozoic is argued to be the initial step. This would then be followed by the development of NDH-1 CO2-uptake systems, followed by the development of low-and high-affinity HCO3– transporters. An intriguing question is, were carboxysomes developed first in cyanobacteria, or did they originate by the lateral transfer of pre-existing proteobacterial bacterial microcompartment genes? The potentially late evolution of the CCM genes in cyanobacteria argues for a polyphyletic and separate evolution of CCMs in cyanobacteria, algae, and higher plants.

Cyanobacteria (blue–green algae) have evolved a remarkable environmental adaptation for survival at limiting CO2 concentrations. The adaptation is known as a CO2 concentrating mechanism, and functions to actively transport and accumulate inorganic carbon (Ci; HCO3– and CO2) within the cell. Thereafter, this Ci pool is utilised to provide elevated CO2 concentrations around the primary CO2 fixing enzyme, Rubisco, which is encapsulated in a unique micro-compartment known as the carboxysome. Recently, significant progress has been gained in understanding the different types of Ci transport in cyanobacteria. This semi-review centres on the model cyanobacterium, Synechococcus sp. PCC7942, which possesses at least four distinct modes of Ci uptake when grown under Ci limitation, each possessing a high degree of functional redundancy. The four modes so far identified are: (i) BCT1, an inducible, high affinity HCO3– transporter of the bacterial ATP binding cassette transporter family, encoded by cmpABCD; (ii) a constitutive, Na+-dependent HCO3– transport system that can be allosterically activated (possibly by phosphorylation) in as little as 10 min; (iii) and (iv) two CO2 uptake systems, one constitutive and the other inducible, based on specialised forms of thylakoid-based, type 1, NAD(P)H dehydrogenase complexes (NDH-1). Here, we forward a speculative model that proposes that two unique proteins, ChpX and ChpY, possess CO2 hydration activity in the light, and when coupled to photosynthetic electron transport through the two specialised NDH-1 complexes, result in net hydration of CO2 to HCO3– as a crucial component of the CO2 uptake process.

Membraneless organelles containing the enzyme Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) are a common feature of organisms utilizing CO2 concentrating mechanisms (CCMs) to enhance photosynthetic carbon acquisition. In cyanobacteria and proteobacteria, the Rubisco condensate is encapsulated in a proteinaceous shell, collectively termed a carboxysome, while some algae and hornworts have evolved Rubisco condensates known as pyrenoids. In both cases, CO2 fixation is enhanced compared with the free enzyme. Previous mathematical models have attributed the improved function of carboxysomes to the generation of elevated CO2 within the organelle via a co-localized carbonic anhydrase (CA), and inwardly diffusing HCO3-which has accumulated in the cytoplasm via dedicated transporters. Here we present a novel concept in which we consider the net of two protons produced in every Rubisco carboxylase reaction. We evaluate this in a reaction-diffusion, compartment model to investigate functional advantages these protons may provide Rubisco condensates and carboxysomes, prior to the evolution of HCO3-accumulation. Our model highlights that diffusional resistance to reaction species within a condensate allows Rubisco-derived protons to drive the conversion of HCO3-to CO2 via co-localized CA, enhancing both condensate [CO2] and Rubisco rate. Protonation of Rubisco substrate (RuBP) and product (PGA) plays an important role in modulating internal pH and CO2 generation. Application of the model to putative evolutionary ancestors, prior to contemporary cellular HCO3-accumulation, revealed photosynthetic enhancements along a logical sequence of advancements, via Rubisco condensation, to fully-formed carboxysomes. Our model suggests that evolution of Rubisco condensation could be favored under low CO2 and low light environments.

Here, we report the complete genome sequences of 12 Mycoplasma bovis isolates cultured from Canadian bison and 4 cultured from Canadian cattle. The sequences are of value for understanding the phylogenetic relationship between cattle and bison isolates and will aid in elucidating the genetic basis for virulence and host specificity.

The ongoing epidemic of chronic wasting disease (CWD) within cervid populations indicates the need for novel approaches for disease management. A vaccine that either reduces susceptibility to infection or reduces shedding of prions by infected animals, or a combination of both, could be of benefit for disease control. The development of such a vaccine is challenged by the unique nature of prion diseases and the requirement for formulation and delivery in an oral format for application in wildlife settings. To address the unique nature of prions, our group targets epitopes, termed disease specific epitopes (DSEs), whose exposure for antibody binding depends on disease-associated misfolding of PrPC into PrPSc. Here, a DSE corresponding to the rigid loop (RL) region, which was immunogenic following parenteral vaccination, was translated into an oral vaccine. This vaccine consists of a replication-incompetent human adenovirus expressing a truncated rabies glycoprotein G recombinant fusion with the RL ...

Mycoplasma bovis is emerging as an important pathogen of farmed bison in North America and is associated with high morbidity and mortality in affected herds. We developed an in-house ELISA to detect antibodies against Mycoplasma spp. in bison sera. The aims of the study were to estimate the seroprevalence against Mycoplasma spp. in bison herds with or without past history of M. bovis–associated disease, and to determine potential risk factors for seropositivity to Mycoplasma spp. in farmed bison in western Canada. A total of 858 serum samples were collected from bison >1 y of age from 19 bison herds. The individual and herd-level seroprevalence of Mycoplasma spp. was 12% and 79%, respectively. The proportion of seropositive animals was 0–41% and 0–9% for herds with or without a history of M. bovis–associated disease, respectively. Mycoplasma spp. appear to be widespread in bison in Manitoba, Saskatchewan, and Alberta. Eight of 11 herds with no history of M. bovis–associated disease were seropositive for Mycoplasma spp., which suggests that bison can be subclinically infected with Mycoplasma spp., or that infection may be underdiagnosed. Although not specific to M. bovis, the in-house ELISA developed to detect antibodies against Mycoplasma spp. may prove to be a valuable herd-level screening tool, providing insight needed for the development of appropriate prevention and control measures for Mycoplasma-related disease in bison herds.

A multi-allelic genetic marker resource for future genetics studies in mānuka (Leptospermum scoparium) and its close relatives has been developed by mining simple sequence repeats (SSR) from a preliminary draft genome assembly of L. scoparium ‘Crimson Glory’. In total, 469 and 169 trinucleotide and dinucleotide repeats, respectively, were detected in a set of 89,619 predicted gene models. Polymerase chain reaction primer pairs were designed for 32 loci exhibiting high sequence similarity to Eucalyptus grandis. These 32 primer pairs were screened over a population of mānuka and 3 accessions of sympatric, related kānuka (genus Kunzea). The 15 polymorphic SSR markers that were successfully developed are now available for genetic analysis in Myrtaceae.

Mott cells are atypical plasmacytes recognized microscopically by endoplasmic reticulum (ER) distensions (Russell bodies) a result of retained secretory product (antibody). Originally associated with parasitism, they are observed in a broad spectrum of immunopathology, sometimes involving hypergammaglobulinemia. Few descriptions of Mott cells appear in avian literature. The purpose of the manuscript is to provide examples identified by light microscopy in three poultry species. Transmission electron micrographs (TEM) of plasmacytes from the turkey oviduct mucosa are included for comparison with Mott cell light microscopic images. Wright's stained blood and bone marrow from commercial and specific pathogen free (SPF) chickens, ducks, and turkeys are the sources. Mott cell positive samples commonly occurred with leukocytosis or leukemoid reactions, polymicrobial bacteremia, and fungemia. Atypical granulocytes and leukocytes regularly accompanied Mott cells. It is proposed that circulating Mott cells are “sentinels” indicative of stress, dyscrasia, and pathology. Moreover, Mott cells, like other atypia, complicate the interpretation of simple heterophil/lymphocyte (H/L) ratios. As Mott cells are defective plasmacytes these observations address hematology, immunology, pathology, and welfare issues.

Reef islands are at the forefront of concern for future accelerating sea-level rise since their low-lying and isolated nature puts them at higher risk of marine inundation compared to continental coastlines. However, the perceived threat of complete submersion as implied by projected future sea-level rise and current island elevations do not consider the morphologically resilient nature of reef island systems. In particular, the role of sediment supply in the resilience of these islands is still relatively poorly studied. This study presents detailed descriptions of the sedimentary characteristics and stratigraphy of two lagoonal platform islands in Huvadhoo Atoll, Maldives, that formed during periods of Holocene sea-level rise. Island subsurface stratigraphy was reconstructed by analysing the skeletal composition and textural properties of 306 sediment samples from 37 cores extracted across the islands. Island sediments were dominated by coral sands with varied proportions of secondary constituents (molluscs, Halimeda, foraminifera, and crustose coralline algae). Downcore variations in composition show that the proportion of coral sands decrease with depth and the proportion of molluscs and Halimeda increase with depth (with the exception of cores that terminated on lagoon infill). The increased proportion of Halimeda and molluscs in these early island deposits may have resulted from the catch-up growth strategy of the reef during the mid-Holocene highstand as both organisms have high turnover rates and directly contribute to sediment production after death. The sedimentological response of increased Halimeda and molluscs highlights the resilient and dynamic nature of reef islands and the ability of reefs to adjust ecologically to changing sea levels.Recorded Presentation from the vICCE (YouTube Link): https://youtu.be/gy3zhqocMGw

Litter decomposition is a key process for ecosystem fertility and carbon (C) balance, key uncertainties remain about how this fundamental process is affected by microbial community composition. Evidence is growing that plant litter generally decays fastest at the site from which it was derived, owing to the presence of specialized microbial communities that can decompose specific types of litter. The objectives were to determine the impact of sites on litter decomposition and to examine the relationships among microbial community composition, litter chemistry, and decomposition rates of coniferous Cunninghamia lanceolata litter of higher lignin content and broadleaved Mytilaria laosensis litter of lower lignin content at different stages of decomposition under plantations of the respective species. The study was conducted for 16 months using a randomized split-plot design experiment with four replications of all combinations of treatments, the treatments being litter type and site (plantation species). The results showed that decomposition rates were the same for all combinations of amendments and sites, meaning that both sites had microbial communities equally capable or adapted to decompose plant substrates it had not previously encountered, despite marked differences in soil microbial communities between sites and the chemistry of the two litter types. Initial M. laosensis litter was of lower lignin content and C:N ratio and decomposed faster in the first 8 months than C. lanceolata litter under either M. laosensis or C. lanceolata forest. Litter decomposition was significantly slower in the environment from which it was derived between month 8 and month 16. This could be attributed to the exceptionally poor decomposition of M. laosensis litter which was significantly higher lingnin content at month 8 under M. laosensis than under C. lanceolata due to the impact of site on preferential degradation of litter C. Decomposers under C. lanceolata forest were more efficient in degrading alkyl C and/or less efficient in degrading O-alkyl C than those under M. laosensis forest during the experimental period, which might be related to the microbial community composition in the decomposing litter. Our study clearly showed interactions between changing litter chemistry and litter microbial communities and their impacts on litter decomposition. Site was not important in impacting decomposition rates, but played an important role in the preferential degradation of C components. However, further studies are needed to examine the conditions in forests where more rapid litter decomposition beneath the parent species than another species is considered to be common, in order to improve our ability to model decomposition globally.

Alfaxalone is a neurosteroid that interacts with gamma-aminobutyric type A receptors to produce central nervous system depression and muscle relaxation. The effects of alfaxalone vary from sedation to general anesthesia. Alfaxalone is synergistic with other tranquilizers and sedatives and therefore has the potential to improve existing alpha-2 adrenergic agonist-based combinations used for wildlife immobilization. The objective of this study was to determine the efficacy and cardiopulmonary effects of a medetomidine-azaperone-alfaxalone (MAA) combination in captive white-tailed deer (Odocoileus virginianus). Eight captive white-tailed deer were restrained in a drop-floor chute; hand injected i.m. with 0.15 mg/kg medetomidine, 0.2 mg/kg azaperone, and 0.5 mg/kg alfaxalone; and released into a small enclosure for observation. The deer were maintained in lateral recumbency for a total time from postinjection (PI) of the drug of 60 min. At 60 min PI, atipamezole was administered i.m. at five times the medetomidine dose. Heart rate, respiratory rate, rectal temperature, and direct systolic, mean, and diastolic arterial blood pressures were recorded every 5 min. Arterial blood samples were taken every 15 min for blood gas analysis. Level of sedation and quality of recovery were scored. Statistical analysis was performed using analysis of variance and descriptive statistics with a significance level of P0.05. Induction (time to lateral recumbency, 7.1 ± 2.4 min (mean ± SD) and recovery times (time to standing, 9.1 ± 3.1 min) were comparable to currently used medetomidine-based combinations in white-tailed deer. Major cardiopulmonary effects observed (values reported are 15 min PI of immobilizing drugs) were hypoxemia (PaO2, 54 ± 9 mm Hg), hypoventilation (PaCO2, 55 ± 3 mm Hg), and mixed acid-base disturbances (pH, 7.22 ± 0.04). No adverse effects were observed after recovery from anesthesia. MAA produced a satisfactory level of deep sedation for safe handling and minor procedures in captive white-tailed deer.

Nitrogen (N) exerts strong effects on litter decomposition through altering microbial abundance and community composition. However, the effect of N addition on plant–soil interactions such as home-field advantage (HFA: enhanced decomposition at a home environment compared to a guest environment) in relation to litter decomposition remains unclear. To fill this knowledge gap, we conducted a reciprocal litter transplant plus N addition experiment in Mytilaria laosensis and Cunninghamia lanceolata plantations for two years in subtropical China where anthropogenic N input is amongst the highest in the world. We found positive HFA effects (in which the calculation incorporates litter of both species) with litter mass loss 11.2% faster at home than in the guest environment in the N addition (50 kg N ha−1 yr−1) treatment, but no significant HFA effects were found in the control treatment. The magnitude of the HFA effect on carbon (C) release increased with N addition, while that on N release decreased. The HFA effects on phosphorus, potassium, calcium, sodium, and magnesium release were positive overall, but varied through time and the magnitude of the effects were different among elements. The greater HFA effects in the N addition treatment were associated with greater differences in microbial biomass and community composition between home and guest environments than in the control treatment. Our results indicate that anthropogenic N enrichment could lead to enhanced HFA effects, through modification of microbial communities, and thereby affect C sequestration and N cycling in subtropical forests.

The range of flower colours in Cymbidium hybrida orchid is extensive. All three main pigment groups have been detected, but our focus has been on the anthocyanins, which may be present in all sepals and petals, only in the labellum, or as a colour blush. Since anthocyanin pigments have an important influence on final flower colour, it is important to study how their biosynthesis is regulated. We report here the isolation of a MYB transcription factor (ChMYB1) that is shown to be an anthocyanin-related MYB and likely to be part of a regulatory complex controlling anthocyanin biosynthesis in Cymbidium flowers. There are at least two separate phases of anthocyanin development in Cymbidium flowers. One occurs during flower bud development, and the other occurs in open flowers after pollination. The relatively rapid induction of anthocyanin biosynthesis after anther cap removal was used to monitor expression of the MYB1 gene and the flavonoid biosynthetic gene dihydroflavonol 4-reductase (DFR). The involvement of ethylene in this anthocyanin induction was examined by applying the ethylene action inhibitor, 1-methyl-cyclopropene (1-MCP), and by studying the expression of ethylene biosynthesis genes. Expression of the MYB1 gene was first detected 12 h after pollinia removal, and it peaked after 24 h, while expression of the DFR gene was first detected at 24 h after pollinia removal and it peaked after 48 h. 1-MCP treatment abolished completely both anthocyanin accumulation and the expression of the ChMYB1 and ChDFR genes. The results suggest that ChMYB1 regulates in Cymbidium hybrida flowers the pollination-induced anthocyanin production mediated by ethylene signalling.

Background Many trichostrongylid nematode species are reported to infect bison, some of which are major causes of disase and production loss in North American bison herds. However, there is little information on the species distribution and relative abundance of these parasites in either commercial or conservation herds. This is largely because trichostrongylid nematode species cannot be distinguished by visual microscopic examination of eggs present in feces. Consequently, we have applied ITS2 rDNA nemabiome metabarcoding to describe the trichostrongyle parasite species diversity in 58 bison production groups derived from 38 commercial North American plains bison (Bison bison bison) herds from across western Canada, and two bison conservation herds located in Elk Island National Park (EINP) [plains bison and wood bison (Bison bison athabascae)] and one in Grasslands National Park (GNP) (plains bison). Results We report much higher infection intensities and parasite species diversity in commercial bison herds than previously reported in beef cattle herds grazing similar latitudes. Predominant trichostrongyle parasite species in western Canadian commercial bison herds are those commonly associated with Canadian cattle, with Ostertagia ostertagi being the most abundant followed by Cooperia oncophora. Combined with high fecal egg counts in many herds, this is consistent with significant clinical and production-limiting gastrointestinal parasitism in western Canadian bison herds. However, Haemonchus placei was the most abundant species in five of the production groups. This is both surprising and important, as this highly pathogenic blood-feeding parasite has not been reported at such abundance, in any livestock species, at such northerly latitudes. The presence of Trichostrongylus axei as the most abundant parasite in four herds is also unusual, relative to cattle. There were striking differences in parasite communities between the EINP and commercial bison herds. Most notably, Orloffia bisonis was the predominant species in the wood bison herd despite being found at only low levels in all other herds surveyed. Conclusions This study represents the most comprehensive description of parasite communities in North American bison to date and illustrates the power of deep amplicon sequencing as a tool to study species diversity in gastrointestinal nematode communities. Electronic supplementary material The online version of this article (10.1186/s13071-018-2880-y) contains supplementary material, which is available to authorized users.

Improvement of yield potential in global food crops has hit a breeding road-block which could be solved through genetic engineering. One promising strategy is the introduction of cyanobacterial CO2-concentrating-mechanisms (CCMs) into plant chloroplasts to enhance photosynthesis and yield. CCMs actively accumulate CO2 within specialized microcompartments called carboxysomes, where the enzyme Rubisco rapidly fixes CO2. Here we replaced the endogenous Rubisco large subunit gene with cyanobacterial Form-1A Rubisco and key structural proteins of α- carboxysomes in tobacco chloroplasts. Transformed plants produced purifiable carboxysomes, isometric with those of the source organism Cyanobium marinum. Carboxysomes encapsulated the introduced Rubisco and enabled autotrophic growth at elevated CO2. Our results demonstrate the formation of α-carboxysomes from a minimal gene set, and that cyanobacterial Form-1A Rubisco supports C3 photosynthesis at high CO2. This major advance informs the step-wise construction of fully-functional α-carboxysomes in chloroplasts which, along with other CCM components, are expected to improve crop plant performance.

The Md - MYB10 R6 gene from apple is capable of self-regulating in heterologous host species and enhancing anthocyanin pigmentation, but the activity of MYB10 is dependent on endogenous protein partners. Coloured foliage due to anthocyanin pigments (bronze/red/black) is an attractive trait that is often lacking in many bedding, ornamental and horticultural plants. Apples (Malus × domestica) containing an allelic variant of the anthocyanin regulator, Md-MYB10 R6 , are highly pigmented throughout the plant, due to autoregulation by MYB10 upon its own promoter. We investigated whether Md-MYB10 R6 from apple is capable of functioning within the heterologous host Petunia hybrida to generate plants with novel pigmentation patterns. The Md-MYB10 R6 transgene (MYB10–R6 pro :MYB10:MYB10 term ) activated anthocyanin synthesis when transiently expressed in Antirrhinum rosea dorsea petals and petunia leaf discs. Stable transgenic petunias containing Md-MYB10 R6 lacked foliar pigmentation but had coloured flowers, complementing the an2 phenotype of ‘Mitchell’ petunia. The absence of foliar pigmentation was due to the failure of the Md-MYB10 R6 gene to self-activate in vegetative tissues, suggesting that additional protein partners are required for Md-MYB10 to activate target genes in this heterologous system. In petunia flowers, where endogenous components including MYB-bHLH-WDR (MBW) proteins were present, expression of the Md-MYB10 R6 promoter was initiated, allowing auto-regulation to occur and activating anthocyanin production. Md-MYB10 is capable of operating within the petunia MBW gene regulation network that controls the expression of the anthocyanin biosynthesis genes, AN1 (bHLH) and MYBx (R3-MYB repressor) in petals.

Located at the anterior end of the turkey hen's vagina are numerous discrete tubular invaginations of the surface epithelium, collectively referred to as the sperm storage tubules (SSTs). After mating or artificial insemination, sperm ascend the vagina, enter the SSTs, and over the ensuing days and weeks, gradually exit the SSTs and are transported to the anterior end of the oviduct to fertilize a daily succession of ova. Little is known regarding the cellular and molecular mechanisms responsible for sperm subsistence in the lumen of the SST. In this study, the origin of microvillus blebs (MvBs) on the apical tips of SST epithelial cells was examined, and their possible role in sperm survival was discussed. Regardless, if sperm are present or not, transmission electron microscopy revealed two types of microvilli differentiated by the presence or absence of pleomorphic unilaminar MvBs localized to their apical tips. Although some MvBs appeared to be discharging their contents into the SST lumen, others appeared to have pinched off the microvillus stem. When SSTs contained clusters of densely packed sperm, the sperm heads of those sperm adjacent to the SST epithelial cell surface were surrounded by the microvilli. Associated with the plasmalemma of sperm throughout the SST lumina were membrane fragments and small vesicles (30-130 nm in diameter), some of which appeared to have fused with sperm. It is concluded that the MvBs are a form of shedding vesicle released from the SST epithelial cell microvilli by apocrine secretion. On the basis of observations described herein and those of other authors, it is suggested that the MvBs contribute to sustained sperm storage in the SSTs by (1) supplying metabolic substrates used by resident sperm, (2) serving as fusogenic vehicles providing exogenous macromolecules that reversibly suppress sperm functions associated with fertilization (decapacitation?) and stabilize the sperm plasmalemma, and (3) acting as transport vesicles actively transporting fluid from the SST epithelial cells to the SST lumen.

This study aimed to determine the influence of tree species on soil microbial community structure. We conducted a litter and root manipulation and a short-term nitrogen (N) addition experiment in 19-year-old broadleaf Mytilaria laosensis (Hamamelidaceae) and coniferous Chinese fir (Cunninghamia lanceolata) plantations in subtropical China. Phospholipid fatty acid (PLFA) analysis was used to examine treatment effects on soil microbial community structure. Redundancy analysis (RDA) was performed to determine the relationships between individual PLFAs and soil properties (soil pH, carbon (C) and N concentration and C:N ratio). Soil C:N ratio was significantly greater in M. laosensis (17.9) than in C. lanceolata (16.2). Soil C:N ratio was the key factor affecting the soil microbial community regardless of tree species and the litter, root and N treatments at our study site. The fungal biomarkers, 18:1ω9 and 18:2ω6,9 were significantly and positively related to soil C:N ratio and the abundance of bacterial lipid biomarkers was negatively related to soil C:N ratio. N addition for 8 months did not change the biomass and structure of the microbial community in M. laosensis and C. lanceolata soils. Soil nutrient availability before N addition was an important factor in determining the effect of N fertilization on soil microbial biomass and activity. PLFA analysis showed that root exclusion significantly decreased the abundance of the fungal biomarkers and increased the abundance of the Gram-positive bacteria. Rootless plots had a relatively lower Gram-positive to Gram-negative bacteria ratio and a higher fungi to bacteria ratio compared to the plots with roots under both M. laosensis and C. lanceolata. The response of arbuscular mycorrhizal fungi (16:1ω5) to root exclusion was species-specific. These observations suggest that soil C:N ratio was an important factor in influencing soil microbial community structure. Further studies are required to confirm the long-term effect of tree species on soil microbial community structure.

This chapter provides a brief overview to the reader a fundamental understanding of the anatomy and physiology of reproduction in birds, with a strong emphasis on poultry. As with all vertebrates, the reproductive system in male birds consist of paired testes that produce sperm and a duct system, in birds referred to as the excurrent ducts, that transports the sperm to a copulatory apparatus found on the ventral floor of the cloaca for transfer to the female at copulation. The cloaca serves as the common opening for the deposition of excretory and digestive wastes and the reproductive tracts in both the male and female birds. With the possible exception of the foam gland found in the roof of the quail proctodeum, there are no accessory sex glands associated with the reproductive tract of male birds. However, at the time of ejaculation poultry semen is diluted with a variable volume of transparent-fluid.

Photorespiration is an energy-intensive process that recycles 2-phosphoglycolate, a toxic product of the Rubisco oxygenation reaction. The photorespiratory pathway is highly compartmentalized, involving the chloroplast, peroxisome, cytosol, and mitochondria. Though the soluble enzymes involved in photorespiration are well characterized, very few membrane transporters involved in photorespiration have been identified to date. In this work, Arabidopsis thaliana plants containing a T-DNA disruption of the bile acid sodium symporter BASS6 show decreased photosynthesis and slower growth under ambient, but not elevated CO2. Exogenous expression of BASS6 complemented this photorespiration mutant phenotype. In addition, metabolite analysis and genetic complementation of glycolate transport in yeast showed that BASS6 was capable of glycolate transport. This is consistent with its involvement in the photorespiratory export of glycolate from Arabidopsis chloroplasts. An Arabidopsis double knockout line of both BASS6 and the glycolate/glycerate transporter PLGG1 (bass6, plgg1) showed an additive growth defect, an increase in glycolate accumulation, and reductions in photosynthetic rates compared with either single mutant. Our data indicate that BASS6 and PLGG1 partner in glycolate export from the chloroplast, whereas PLGG1 alone accounts for the import of glycerate. BASS6 and PLGG1 therefore balance the export of two glycolate molecules with the import of one glycerate molecule during photorespiration.

Mitochondrial respiration in the dark (R dark) is a critical plant physiological process, and hence a reliable, efficient and high-throughput method of measuring variation in rates of R dark is essential for agronomic and ecological studies. However, currently methods used to measure R dark in plant tissues are typically low throughput. We assessed a high-throughput automated fluorophore system of detecting multiple O2 consumption rates. The fluorophore technique was compared with O2-electrodes, infrared gas analysers (IRGA), and membrane inlet mass spectrometry, to determine accuracy and speed of detecting respiratory fluxes. The high-throughput fluorophore system provided stable measurements of R dark in detached leaf and root tissues over many hours. High-throughput potential was evident in that the fluorophore system was 10 to 26-fold faster per sample measurement than other conventional methods. The versatility of the technique was evident in its enabling: (1) rapid screening of R dark in 138 genotypes of wheat; and, (2) quantification of rarely-assessed whole-plant R dark through dissection and simultaneous measurements of above- and below-ground organs. Variation in absolute R dark was observed between techniques, likely due to variation in sample conditions (i.e. liquid vs. gas-phase, open vs. closed systems), indicating that comparisons between studies using different measuring apparatus may not be feasible. However, the high-throughput protocol we present provided similar values of R dark to the most commonly used IRGA instrument currently employed by plant scientists. Together with the greater than tenfold increase in sample processing speed, we conclude that the high-throughput protocol enables reliable, stable and reproducible measurements of R dark on multiple samples simultaneously, irrespective of plant or tissue type.

Improving global yields of agricultural crops is a complex challenge with evidence indicating benefits in productivity are achieved by enhancing photosynthetic carbon assimilation. Towards improving rates of CO2 capture within leaf chloroplasts, this study shows the versatility of plastome transformation for expressing the Synechococcus PCC7002 BicA bicarbonate transporter within tobacco plastids. Fractionation of chloroplast membranes from transplastomic tob(BicA) lines showed that ~75% of the BicA localized to the thylakoid membranes and ~25% to the chloroplast envelope. BicA levels were highest in young emerging tob(BicA) leaves (0.12 μmol m(-2), ≈7mg m(-2)) accounting for ~0.1% (w/w) of the leaf protein. In these leaves, the molar amount of BicA was 16-fold lower than the abundant thylakoid photosystem II D1 protein (~1.9 μmol m(-2)) which was comparable to the 9:1 molar ratio of D1:BicA measured in air-grown Synechococcus PCC7002 cells. The BicA produced had no discernible effect on chloroplast ultrastructure, photosynthetic CO2-assimilation rates, carbon isotope discrimination, or growth of the tob(BicA) plants, implying that the bicarbonate transporter had little or no activity. These findings demonstrate the utility of plastome transformation for targeting bicarbonate transporter proteins into the chloroplast membranes without impeding growth or plastid ultrastructure. This study establishes the span of experimental measurements required to verify heterologous bicarbonate transporter function and location in chloroplasts and underscores the need for more detailed understanding of BicA structure and function to identify solutions for enabling its activation and operation in leaf chloroplasts.

The carbon dioxide (CO 2 )-concentrating mechanism of cyanobacteria is characterized by the occurrence of Rubisco-containing microcompartments called carboxysomes within cells. The encapsulation of Rubisco allows for high-CO 2 concentrations at the site of fixation, providing an advantage in low-CO 2 environments. Cyanobacteria with Form-IA Rubisco contain α-carboxysomes, and cyanobacteria with Form-IB Rubisco contain β-carboxysomes. The two carboxysome types have arisen through convergent evolution, and α-cyanobacteria and β-cyanobacteria occupy different ecological niches. Here, we present, to our knowledge, the first direct comparison of the carboxysome function from α-cyanobacteria ( Cyanobium spp. PCC7001) and β-cyanobacteria ( Synechococcus spp. PCC7942) with similar inorganic carbon (C i ; as CO 2 and HCO 3 − ) transporter systems. Despite evolutionary and structural differences between α-carboxysomes and β-carboxysomes, we found that the two strains are remarkably similar in many physiological parameters, particularly the response of photosynthesis to light and external C i and their modulation of internal ribulose-1,5-bisphosphate, phosphoglycerate, and C i pools when grown under comparable conditions. In addition, the different Rubisco forms present in each carboxysome had almost identical kinetic parameters. The conclusions indicate that the possession of different carboxysome types does not significantly influence the physiological function of these species and that similar carboxysome function may be possessed by each carboxysome type. Interestingly, both carboxysome types showed a response to cytosolic C i , which is of higher affinity than predicted by current models, being saturated by 5 to 15 mmC i . This finding has bearing on the viability of transplanting functional carboxysomes into the C 3 chloroplast.

Plants require sophisticated regulatory mechanisms to ensure the degree of anthocyanin pigmentation is appropriate to myriad developmental and environmental signals. Central to this process are the activity of MYB-bHLH-WD repeat (MBW) complexes that regulate the transcription of anthocyanin genes. In this study, the gene regulatory network that regulates anthocyanin synthesis in petunia (Petunia hybrida) has been characterized. Genetic and molecular evidence show that the R2R3-MYB, MYB27, is an anthocyanin repressor that functions as part of the MBW complex and represses transcription through its C-terminal EAR motif. MYB27 targets both the anthocyanin pathway genes and basic-helix-loop-helix (bHLH) ANTHOCYANIN1 (AN1), itself an essential component of the MBW activation complex for pigmentation. Other features of the regulatory network identified include inhibition of AN1 activity by the competitive R3-MYB repressor MYBx and the activation of AN1, MYB27, and MYBx by the MBW activation complex, providing for both reinforcement and feedback regulation. We also demonstrate the intercellular movement of the WDR protein (AN11) and R3-repressor (MYBx), which may facilitate anthocyanin pigment pattern formation. The fundamental features of this regulatory network in the Asterid model of petunia are similar to those in the Rosid model of Arabidopsis thaliana and are thus likely to be widespread in the Eudicots.

The objective of the study was to establish an effective ovarian superstimulatory protocol and subsequently obtain oocytes from bison by transvaginal ultrasound-guided follicular aspiration. Two experiments involving 22 wood bison were done during the breeding season (September to December). In experiment 1, the bison were given a luteolytic dose of prostaglandin (Day 0) and underwent follicular ablation (Day 8) to induce ovarian synchrony. Synchronized bison were then assigned randomly to two groups (n = 11 per group) and given either 200 mg FSH diluted in saline sc, or 200 mg FSH diluted in a proprietary slow-release formulation (SRF) im on Days 9 and 11. Prostaglandin was given to both groups on Day 11 followed by 25 mg LH on Day 13. Oocytes were collected by transvaginal ultrasound-guided aspiration of follicles ≥5 mm on Day 14. In experiment 2, bison were synchronized as in experiment 1 and assigned randomly to one of two groups (n = 11 per group) and given either a single dose of 2500 IU eCG im on Day 9, or 200 mg FSH sc on Days 9 and 11. Prostaglandin was given to both groups on Day 11, and LH (25 mg) was given on Day 13. Oocyte collection was done as described in experiment 1. Cumulus-oocyte-complexes (COC) were classified according to morphologic characteristics. In experiment 1, more follicles ≥5 mm were detected on Day 14 in bison treated with FSH versus eCG (12.2 ± 1.73 vs. 5.8 ± 0.52; P < 0.05), and more COC were collected from FSH-treated animals (7.2 ± 1.41 vs. 3.4 ± 0.62; P < 0.05). In experiment 2, the FSH-saline and FSH-SRF groups had a similar number (mean value ± standard error of the mean) of follicles ≥5 mm on Day 14 (12.4 ± 1.49 vs. 13.8 ± 1.24, respectively) and a similar number of COC were collected (6.5 ± 1.13 vs. 6.3 ± 0.96, respectively). The proportion of COC collected per follicle aspirated and the percentage of compact, expanded, and denuded oocytes did not differ between groups in either experiment 1 or 2. In summary, a two-dose regimen of FSH diluted in saline and given sc or in a SRF and given im induced a similar ovarian response in wood bison, whereas a single dose of eCG resulted in a significantly lower ovarian response. Overall, COC were collected from 55% of follicles after transvaginal, ultrasound-guided needle aspiration in wood bison.

Experiments were done to determine if inclusion of eCG and progesterone in the superstimulation protocol will increase the ovarian response and embryo production in wood bison, and to provide preliminary information regarding the effect of season. In Experiment 1 (anovulatory season), bison (n=26) were synchronized by follicular ablation (Day -1) and given FSH on Days 0 and 2, and assigned to 3 groups: Progesterone (Days 0-4), eCG (Day 3), or progesterone+eCG. On Day 5, bison were given hCG and inseminated 12 and 24h later. Ova/embryos were collected 8days after hCG. In Experiment 2 (ovulatory season), bison (n=24) were synchronized and assigned randomly to two groups in which superstimulation was induced with FSH, either with or without eCG, as in Experiment 1. No differences among groups were found in ovarian response or embryo production in either experiment. The follicular count at wave emergence was positively correlated with the number of large follicles at the end of superstimulation in all groups. A significantly greater number of follicles present at wave emergence in the anovulatory vs. ovulatory season was associated with a greater number of CL at the time of embryo collection, but only half the number of freezable embryos. In conclusion, the number of transferable embryos collected (1-2/bison) was higher than in any previous report, but was not attributable to the inclusion of eCG or progesterone in the superovulatory protocol. The apparent effect of season on oocyte competence, and not superovulatory response, is worthy of further investigation.

The identification, enrichment and subsequent isolation of spermatogonial stem cells (SSCs) are integral to the success of SCC transplants between fertile donor and sterilized recipient males. In birds generally and particularly in chicken, SSC-specific has yet to be identified. The receptor for glial cell-derived neurotrophic factor (GDNF), i.e. GDNF family receptor alpha-1 (GFRα1), has been identified as a potential marker for different mouse spermatogonial subtypes. In the present study, we characterized the chicken cGFRα1 receptor and compared its predicted amino-acid sequence with mouse, rat and human GFRα1 proteins. Using specific polyclonal mouse anti-cGFRα1 serum, a total of 2.8% cells were recognized as cGFRα1-positive among isolated testicular cells recovered from sexually mature cockerels. The percentages of cGFRα1-positive testicular cells with haploid, diploid, tetraploid and SP DNA content were 1.6%, 2.5%, 39.3% and 76.8%, respectively. The presence of cGFRα1 protein on the surfaces of all cells of the seminiferous epithelium was confirmed by immunocytochemical and immunohistochemical analyses. Tissue specificity of cGFRα1 mRNA expression was significantly higher in adult testes compared to brain tissue which itself was several times higher than tissues prepared from the spleen, liver and heart. No expression was observed in muscular tissue. At last, we demonstrated the successful repopulation of sterilized recipient's testes with transplanted cGFRα1-positive donor testicular cells. Recipient males subsequently produced functional heterologous spermatozoa capable of fertilizing an ovum and obtaining chicks with donor cell genotypes.

Plant nutrient uptake from coarse soil (2–4 mm diameter) has been demonstrated for only a limited number of nutrients, and the nutritional contribution of coarse soil when present with fine soil (material

SUMMARY Cyanobacteria are the globally dominant photoautotrophic lineage. Their success is dependent on a set of adaptations collectively termed the CO 2 -concentrating mechanism (CCM). The purpose of the CCM is to support effective CO 2 fixation by enhancing the chemical conditions in the vicinity of the primary CO 2 -fixing enzyme, d -ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO), to promote the carboxylase reaction and suppress the oxygenase reaction. In cyanobacteria and some proteobacteria, this is achieved by encapsulation of RubisCO within carboxysomes, which are examples of a group of proteinaceous bodies called bacterial microcompartments. Carboxysomes encapsulate the CO 2 -fixing enzyme within the selectively permeable protein shell and simultaneously encapsulate a carbonic anhydrase enzyme for CO 2 supply from a cytoplasmic bicarbonate pool. These bodies appear to have arisen twice and undergone a process of convergent evolution. While the gross structures of all known carboxysomes are ostensibly very similar, with shared gross features such as a selectively permeable shell layer, each type of carboxysome encapsulates a phyletically distinct form of RubisCO enzyme. Furthermore, the specific proteins forming structures such as the protein shell or the inner RubisCO matrix are not identical between carboxysome types. Each type has evolutionarily distinct forms of the same proteins, as well as proteins that are entirely unrelated to one another. In light of recent developments in the study of carboxysome structure and function, we present this review to summarize the knowledge of the structure and function of both types of carboxysome. We also endeavor to cast light on differing evolutionary trajectories which may have led to the differences observed in extant carboxysomes.

Biochemical models are used to predict and understand the response of photosynthesis to rising temperatures and CO2 partial pressures. These models require the temperature dependency of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) kinetics and mesophyll conductance to CO2 (g(m)). However, it is not known how the temperature response of Rubisco kinetics differs between species, and comprehensive in vivo Rubisco kinetics that include gm have only been determined in the warm-adapted Nicotiana tabacum. Here, we measured the temperature response of Rubisco kinetics and gm in N. tabacum and the cold-adapted Arabidopsis thaliana using gas exchange and (13)CO2 isotopic discrimination on plants with genetically reduced levels of Rubisco. While the individual Rubisco kinetic parameters in N. tabacum and A. thaliana were similar across temperatures, they collectively resulted in significantly different modelled rates of photosynthesis. Additionally, gm increased with temperature in N. tabacum but not in A. thaliana. These findings highlight the importance of considering species-dependent differences in Rubisco kinetics and gm when modelling the temperature response of photosynthesis.

This study investigated the genotypic variation in foliar nutrient concentrations, isotopic signature (δ13C), and chlorophyll fluorescence (Fv/Fm) and tree growth of 40 radiata pine clones grown on a New Zealand serpentine soil, and the relationships between growth and physiological traits of these clones from improved and unimproved groups. Genotypic variation in growth and physiological traits existed within (i.e., clonal) and between groups, with larger variation among clones. The clonal repeatabilities were greater for foliar nitrogen (N), calcium (Ca), magnesium (Mg), boron (B) concentrations, δ13C, and Ca : Mg ratio (0.35–0.64) than for growth traits (0.14–0.27) and other physiological traits (0.08–0.24). Significant phenotypic correlations were found between growth traits and foliar phosphorus (P), potassium (K), sulfur (S), iron (Fe), and K : Mg and Ca : Mg ratios and Fv/Fm (positive), and foliar Mg (negative). This study indicates that the trees on this serpentine soil generally suffered from multiple nutrient deficiencies and imbalances and the clonal variation in growth performance was more related to their capabilities of acclimation to nutrient than water stresses. Overall, the clones that absorbed more P, K, S, and Fe and less Mg from the soil grew better on this serpentine soil. For unimproved clones, the most limiting nutrients for tree growth were foliar K and Fe, while for improved clones it was foliar K.

The objective was to evaluate the efficacy of three previously unreported ovarian superovulatory treatment protocols in wapiti. Protocols were initiated specifically at the time of ovarian follicular wave emergence, and intended to enable determination of the effects of frequency of treatment (i.e., animal handling) and LH supplementation on ovarian response. Thirteen parous wapiti hinds, 2 to 4 y of age, were used late in the anovulatory season (July). The ovaries were examined daily by transrectal ultrasonography. Hinds were given 5 mg estradiol 17-β im (day of treatment designated as Day 0) to induce a new wave of ovarian follicular development. On the expected day of wave emergence (Day 3), hinds were assigned randomly to three treatment groups and given: (1) 100 mg FSH im once a day for 4 days (N = 5); (2) 200 mg FSH sc on Day 3 and Day 5 (N = 4); or (3) 200 mg FSH plus 2.5 mg LH sc on Day 3 and Day 5 (N = 4). All hinds were given 10 mg LH im on Day 6 to induce ovulation. The mean (± SEM) number of ovulations per animal in the respective groups was 6.2 ± 2.0, 15.5 ± 5.9, and 14.8 ± 2.7. In conclusion, the technique of inducing follicular wave emergence to initiate superovulatory treatment at the time of wave emergence was effective in wapiti during the anovulatory season. The most efficient and effective method of ovarian superovulation in this study involved administration of estradiol 17-β on Day 0, followed by 200 mg FSH sc on Days 3 and 5, and induction of ovulation (10 mg of LH) on the evening of Day 6. Compared with conventional methods that require 14 days and handling the hinds six times, the protocol used herein reduced the treatment period to 8 days and the number of animal handlings to four.

Crop yields need to nearly double over the next 35 years to keep pace with projected population growth. Improving photosynthesis, via a range of genetic engineering strategies, has been identified as a promising target for crop improvement with regard to increased photosynthetic yield and better water-use efficiency (WUE). One approach is based on integrating components of the highly efficient CO(2)-concentrating mechanism (CCM) present in cyanobacteria (blue-green algae) into the chloroplasts of key C(3) crop plants, particularly wheat and rice. Four progressive phases towards engineering components of the cyanobacterial CCM into C(3) species can be envisaged. The first phase (1a), and simplest, is to consider the transplantation of cyanobacterial bicarbonate transporters to C(3) chloroplasts, by host genomic expression and chloroplast targeting, to raise CO(2) levels in the chloroplast and provide a significant improvement in photosynthetic performance. Mathematical modelling indicates that improvements in photosynthesis as high as 28% could be achieved by introducing both of the single-gene, cyanobacterial bicarbonate transporters, known as BicA and SbtA, into C(3) plant chloroplasts. Part of the first phase (1b) includes the more challenging integration of a functional cyanobacterial carboxysome into the chloroplast by chloroplast genome transformation. The later three phases would be progressively more elaborate, taking longer to engineer other functional components of the cyanobacterial CCM into the chloroplast, and targeting photosynthetic and WUE efficiencies typical of C(4) photosynthesis. These later stages would include the addition of NDH-1-type CO(2) pumps and suppression of carbonic anhydrase and C(3) Rubisco in the chloroplast stroma. We include a score card for assessing the success of physiological modifications gained in phase 1a.

Most major crops used for human consumption are C3 plants, which yields are limited by photosynthetic inefficiency. To circumvent this, it has been proposed to implement the cyanobacterial CO2-concentrating mechanism (CCM), principally consisting of bicarbonate transporters and carboxysomes, into plant chloroplasts. As it is currently not possible to recover homoplasmic transplastomic monocots, foreign genes must be introduced in these plants via nuclear transformation. Consequently, it is paramount to ensure that resulting proteins reach the appropriate sub-cellular compartment, which for cyanobacterial transporters BicA and SbtA, is the chloroplast inner-envelope membrane (IEM). At present, targeting signals to redirect large transmembrane proteins from non-chloroplastic organisms to plant chloroplast envelopes are unknown. The goal of this study was to identify such signals, using agrobacteria-mediated transient expression and confocal microscopy to determine the sub-cellular localization of ~37 GFP-tagged chimeras. Initially, fragments of chloroplast proteins known to target soluble cargos to the stroma were tested for their ability to redirect BicA, but they proved ineffective. Next, different N-terminal regions from Arabidopsis IEM transporters were tested. We demonstrated that the N-terminus of AtHP59, AtPLGG1 or AtNTT1 (92-115 amino acids), containing a cleavable chloroplast transit peptide (cTP) and a membrane protein leader (MPL), was sufficient to redirect BicA or SbtA to the chloroplast envelope. This constitutes the first evidence that nuclear-encoded transmembrane proteins from non-chloroplastic organisms can be targeted to the envelope of plant chloroplasts; a finding which represents an important advance in chloroplast engineering by opening up the door to further manipulation of the chloroplastic envelope.

d 13 C, d 15 N. Summary � The impact of long-term nitrogen (N) deposition is under-studied in phosphorus (P)-limited subtropical forests. We exploited historically collected herbarium specimens to investigate potential physiological responses of trees in three subtropical forests representing an urbanto-rural gradient, across which N deposition has probably varied over the past six decades. We measured foliar [N] and [P] and stable carbon (d 13 C), oxygen (d 18 O) and nitrogen (d 15 N) isotopic compositions in tissue from herbarium specimens of plant species collected from 1947 to 2014. � Foliar [N] and N : P increased, and d 15 N and [P] decreased in the two forests close to urban centers. Consistent with recent studies demonstrating that N deposition in the region is 15 N-depleted, these data suggest that the increased foliar [N] and N : P, and decreased [P], may be attributable to atmospheric deposition and associated enhancement of P limitation. � Estimates of intrinsic water use efficiency calculated from foliar d 13 C decreased by c. 30% from the 1950s to 2014, contrasting with multiple studies investigating similar parameters in N-limited forests. This effect may reflect decreased photosynthesis, as suggested by a conceptual model of foliar d 13 C and d 18 O. � Long-term N deposition may exacerbate P limitation and mitigate projected increases in carbon stocks driven by elevated CO2 in forests on P-limited soils.