Morphometric characterization of the airway and vascular systems of the lung of the domestic pig, Sus scrofa: comparison of the airway, arterial and venous systems.

The bronchial system (BS), the pulmonary artery (PA) and the pulmonary vein (PV) of the lung of the domestic pig, Sus scrofa were simultaneously cast with silicone rubber and studied. Asymmetrical dichotomous bifurcation preponderated in the tree-like arrangement of the three conducting systems. Lengths and diameters of the various generations were measured. At the extremities of the BS and the PA, alveoli and blood capillaries related very closely. In the cranial and middle lobes of the right and left lungs, topographically, the PA and the PV closely followed the BS, but in the accessory and the caudal (diaphragmatic) lobes, only the PA accompanied the BS: the PV run intersegmentally. Certain similarities and differences were observed between the diameters and lengths of the various generations of the three conducting systems. The strong correlations between some of the structural parameters indicated a high level of structural optimization. While morphometric variations suggest that the air and the blood flow dynamics may somewhat differ between the three conducting systems, they may also register structural features unique to the lung of the domestic pig, an animal that has been highly genetically exploited for fast growth and now leads an indolent lifestyle in captivity.     

Functional cues in the development of osseous tooth support in the pig, Sus scrofa

Alveolar bone supports teeth during chewing through a ligamentous interface with tooth roots. Although tooth loads are presumed to direct the development and adaptation of these tissues, strain distribution in the alveolar bone at different stages of tooth eruption and periodontal development is unknown. This study investigates the biomechanical effects of tooth loading on developing alveolar bone as a tooth erupts into occlusion. Mandibular segments from miniature pigs, Sus scrofa, containing M₁ either erupting or in functional occlusion, were loaded in compression. Simultaneous recordings were made from rosette strain gages affixed to the lingual alveolar bone and the M₂ crypt. Overall, specimens with erupting M₁s were more deformable than specimens with occluding M₁s (mean stiffness of 246 vs. 944 MPa, respectively, p=0.004). The major difference in alveolar strain between the two stages was in orientation. The vertically applied compressive loads were more directly reflected in the alveolar bone strains of erupting M₁s, than those of occluding M₁s, presumably because of the mediation of a more mature periodontal ligament (PDL) in the latter. The PDL interface between occluding teeth and alveolar bone is likely to stiffen the system, allowing transmission of occlusal loads. Alveolar strains may provide a stimulus for bone growth in the alveolar process and crest.     

Comparison of the digestible energy content of maize, oats and alfalfa between the European wild boar (Sus scrofa L.) and Landrace × Large White pig (Sus scrofa domesticus)

The objective of the current study was to compare the digestible energy (DE) contents of maize, oats and alfalfa meal between European wild boar (Sus scrofa L.) and the domestic pig (S. scrofa domesticus, Landrace × Large White). Six pure wild boar (S. scrofa L.) and six domestic pigs (Landrace × Large White) with liveweights (mean ± S.E.M.) of 26 ± 0.6 and 21 ± 1.1 kg, respectively, were fed diets at a daily level of 0.10 × metabolic body weight (W^0.75). The diets included a base diet and three experimental diets containing 700 g basal diet/kg and 300 g maize, oats or alfalfa meal/kg; all animals received all four diets. Chromic oxide was used as indigestible marker. The animals received each diet for an 8-day period with fecal samples collected on days 6, 7 and 8. The DE content of the maize, oats and alfalfa was calculated for each ingredient and statistically compared between the wild boar and domestic pig. For the maize and oats, there was no significant difference in the DE values between the domestic pig and wild boar. However, the DE value of the alfalfa was greater in the domestic pig (10.56 MJ kg⁻¹ DM) than in the wild boar (8.48 MJ kg⁻¹ DM). For ingredients that contain relatively low concentrations of fibre (such as maize and oats), it appears that DE values determined in the domestic pig can be validly applied for diet formulation for wild boars; however, for ingredients with higher fibre levels, the DE values in wild boar appear to be lower than those in the domestic pig.     

The blood respiratory, haematological, acid-base and ionic status of the Port Jackson shark, Heterodontusportusjacksoni, during recovery from anaesthesia and surgery: a comparison with sampling by direct caudal puncture

The effects of caudal cannulation on the blood physiology of the Port Jackson shark, Heterodontus portusjacksoni, were investigated in sharks given between 4 and 72 h to recover from surgery. Neither the Pa_O2–Pv_O2 difference nor the Ca_O2–Cv_O2 difference of cannulated sharks fluctuated throughout the sampling period. The plasma acidosis exhibited 4 h after surgery was partially compensated after 24 h by a respiratory (hyperventilatory) alkalosis and after 72 h by a marked metabolic alkalosis. Whilst H. portusjacksoni exhibited some cell swelling after surgery the haematological status of cannulated sharks generally varied little throughout the recovery period. In contrast, marked changes in plasma and erythrocyte ion concentrations were indicative of increased branchial and erythrocyte ion permeability. The blood status of H. portusjacksoni given 72 h to recover from surgery was also compared with sharks sampled by caudal puncture. The respiratory and acid-base status of sharks sampled by caudal puncture was comparable to that of cannulated sharks. In contrast, the plasma ion concentrations of the cannulated sharks were markedly elevated and the erythrocyte ion concentrations concomitantly reduced when compared with punctured sharks. The apparent increase in the water and ion permeability of cannulated sharks was reflected by the reduced [Hb] and mean cell haemoglobin concentrations (MCHC). Blood sampling by caudal puncture appeared to reduce the haematological and ionic perturbations that resulted from surgery and thus provided a less invasive and reliable method for obtaining samples from ‘non-disturbed’ elasmobranchs.     

Hunger rapidly overrides the risk of predation in the subtidal scavenger Nassarius siquijorensis (Gastropoda: Nassariidae): an energy budget and a comparison with the intertidal Nassarius festivus in Hong Kong

This study shows that, as with its intertidal counterpart, Nassarius festivus, the rate at which subtidal Nassarius siquijorensis moves towards food bait is similar for starved and well-fed individuals. This study also investigates another facet of nassariid nutrition related to the degree of hunger, i.e. the effect of simulated predation upon a feeding assemblage. Individuals which fed within 7 days, cease feeding and depart palatable food if crushed conspecifics are added. Between 7 and 13 days since its last meal, however, N. siquijorensis will feed when food is available, despite the possibility of predation. For the intertidal N. festivus, the critical time for hunger to override the risk of predation is between 14 and 21 days. The difference between subtidal and intertidal species may be due to a difference, in terms of days, that a meal can provide for their energy expenditure, particularly with regard to respiration. The bigger, subtidal, N. siquijorensis needs to feed more frequently than the smaller, intertidal, N. festivus.     

A systematic study of the development of the airway (bronchial) system of the avian lung from days 3 to 26 of embryogenesis: a transmission electron microscopic study on the domestic fowl, Gallus gallus variant domesticus

In the embryo of the domestic fowl, Gallus gallus variant domesticus, the lung buds become evident on day 3 of development. After fusing on the ventral midline, the single entity divides into left and right primordial lungs that elongate caudally while diverging and shifting towards the dorsolateral aspects of the coelomic cavity. On reaching their definitive topographical locations, the lungs rotate along a longitudinal axis, attach, and begin to slide into the ribs. First appearing as a solid cord of epithelial cells that runs in the proximal-distal axis of the developing lung, progressively, the intrapulmonary primary bronchus begins to canalize. In quick succession, secondary bronchi sprout from it in a craniocaudal sequence and radiate outwards. On reaching the periphery of the lung, parabronchi (tertiary bronchi) bud from the secondary bronchi and project into the surrounding mesenchymal cell mass. The parabronchi canalize, lengthen, increase in diameter, anastomose, and ultimately connect the secondary bronchi. The luminal aspect of the formative parabronchi is initially lined by a composite epithelium of which the peripheral cells attach onto the basement membrane while the apical ones project prominently into the lumen. The epithelium transforms to a simple columnar type in which the cells connect through arm-like extensions and prominently large intercellular spaces form. The atria are conspicuous on day 15, the infundibulae on day 16, and air capillaries on day 18. At hatching (day 21), the air and blood capillaries have anastomosed profusely and the blood-gas barrier become remarkably thin. The lung is well developed and potentially functionally competent at the end of the embryonic life. Thereafter, at least upto day 26, no further consequential structures form. The mechanisms by which the airways in the avian lung develop fundamentally differ from those that occur in the mammalian one. Compared with the blind-ended bronchial system that inaugurates in the mammalian lung, an elaborate, continuous system of air conduits develops in the avian one. Further studies are necessary to underpin the specific molecular factors and genetic processes that direct the morphogenesis of an exceptionally complex and efficient respiratory organ.     

Seasonal changes in photosynthesis, antioxidant systems and ELIP expression in a thermonastic and non-thermonastic Rhododendron species: A comparison of photoprotective strategies in overwintering plants

Leaves of overwintering evergreen rhododendrons are typically exposed to freezing temperatures and high light during winters which can potentially result in photon flux exceeding that required for photochemistry. This excess energy, if not dissipated as heat or fluorescence, may cause photooxidative damage to PSII. The goal of this study is to compare the photoprotection strategies during seasonal cold acclimation (CA) in two Rhododendron species (R. catawbiense Michx. and R. ponticum L.) that are divergent in their leaf freezing tolerance and thermonastic behaviour (temperature-induced leaf movement). R. catawbiense exhibits thermonasty while R. ponticum does not. Differences in leaf freezing tolerance (LT₅₀), photosynthesis, photoinhibition, early light-induced proteins (ELIPs) gene expression, and accumulation of antioxidant metabolites and enzymes during seasonal CA were investigated. During seasonal CA, maximum photosynthetic rate (P_max) and maximum quantum efficiency of PSII (F_v/F_m) were significantly down-regulated. Compared with R. catawbiense, R. ponticum showed less photoinhibition and higher overall accumulation (in magnitude) of antioxidant systems while R. catawbiense exhibited more efficient up-regulation of ELIP expression and antioxidant system (i.e., greater efficiency of increasing this pool in winter months relative to the summer levels). The two species respond differently to winter conditions and have evolved strategies to avoid, reduce and/or tolerate photooxidative stress in winter. These include down-regulation of photosynthesis and up-regulation of ELIPs and antioxidant systems, together with specialized leaf anatomy and thermonasty behaviour.     

The flagellates of the termite Hodotermopsis sjoestedti: Immunological and ultrastructural characterization of four new species in the genera Spirotrichonympha, Spironympha and Microjoenia

Four species of spirotrichonymphids representing three genera Spirotrichonympha, Spironympha and Microjoenia symbiotic in the termite Hodotermopsis sjoestedti, have been studied by light and immunofluorescence microscopy and transmission electron microscopy. The genus Spirotrichonympha represented by Spirotrichonympha cincta n. sp. is characterized by a compound axostyle composed of several fibers or subaxostyles, and this species has peculiar undescribed structures associated with the flagellar lines. The genus Spironympha is characterized by flagellar lines restricted to the anterior area and a simple tubular axostyle limited by 1–3 layers of microtubules. The two new Spironympha species Spironympha obtusa and Spironympha oblonga are distinguished by peculiarities in the flagellar lines and the axostyle, as revealed by immunofluorescence and electron microscopy. The genus Microjoenia represented by Microjoenia minuta n. sp. has radiating flagellar lines and an axostyle with two microtubular rows originating from the pelta-axostyle rows covering the anterior cap. Four new species were named and one was assigned to the SSU rRNA sequences provided by the molecular phylogenetic studies by Ohkuma et al. [2000. Phylogenetic identification of hypermastigotes, Pseudotrichonympha, Spirotrichonympha, Holomastigotes, and parabasalian symbionts in the hindgut of termites. J. Eukaryot. Microbiol. 47, 249–259; 2005. Molecular phylogeny of parabasalids inferred from small subunit rRNA sequences, with emphasis on the Hypermastigea. Mol. Phylogenet. Evol. 35, 646–655].     

Studies on metabolic properties in the Northern Krill, Meganyctiphanes norvegica (Crustacea, Euphausiacea): influence of nutrition and season on pyruvate kinase

The specific activity and the kinetic properties of partly purified pyruvate kinase (PK) (EC 2.7.1.40) from the Northern Krill, Meganyctiphanes norvegica, were investigated in relation to varying food resources. In order to evaluate the effect of starvation on the total energy metabolism, the respiration rates of fed and unfed krill were determined. The FPLC–elution profile of PK displayed two distinct peaks — PK I and II. The first isoform represented 80% of the total PK activity in the organism, and 20% was contributed by the second isoform. PK I was inhibited by ATP but was not influenced by fructose–1,6–bisphosphate (FBP). In contrast, PK II showed ATP inhibition and up to 2.5-fold increased activity by addition of 17 μmol·l⁻¹ FBP. The Michaelis–Menten constants of both isoforms were 2–10-fold higher for ADP than for phosphoenolpyruvate (PEP). Alanine showed no regulatory effect on PK I and II. In specimens starved for 7 days oxygen consumption decreased by 20%. Neither the feeding experiments nor the animals captured in the field during low and high productive seasons indicate that PK properties of M. norvegica are modified in relation to food supply. Accordingly, alternative mechanisms are involved in the depression of the metabolic rate in terms of oxygen consumption.     

Isolation and characterization of the krev-1 gene, a novel member of ras superfamily in Neurospora crassa : involvement in sexual cycle progression

Genes belonging to the ras superfamily encode low-molecular-weight GTP/GDP-binding proteins that are highly conserved in wide variety of organisms. We used the polymerase chain reaction (PCR) to isolate a novel member of the ras superfamily from the filamentous fungus Neurospora crassa and obtained a mammalian Krev-1 homolog. We named the gene krev-1 and analyzed its structure and function. The krev-1 gene encodes a polypeptide of 225 amino acids, which is nearly 60% homologous to the mammalian Krev-1 p21. The krev-1 gene product (KREV1) is functionally analogous to mammalian Krev-1 p21 and Rsr1p/Bud1p, a Krev-1 homolog from the yeast Saccharomyces cerevisiae. GAL1-driven expression of KREV1 in a wild-type yeast strain resulted in a random budding pattern, as did its mammalian counterpart Krev-1 p21. We disrupted the krev-1 gene by RIP (repeat-induced point mutation), but the krev-1 disruptants showed no abnormalities. By in vitro mutagenesis, we constructed several mutant krev-1 genes (G21V, A68T, and D128A) which mimic constitutively active mutants of Ha-ras, and the krev-1 (K25N) mutant which is analogous to a dominant-negative mutant of Ha-ras. Each mutant gene was introduced into the wild-type strain and the phenotypes were analyzed. We could not observe any difference in vegetative growth between these transformants. When each strain was used as the female in mating tests, the development of perithecia from protoperithecia was inhibited in all cases. The results indicate that the krev-1 gene may be involved in sexual cycle progression. Received: 28 January 1997 / Accepted: 3 April 1997     

Cloning and characterization of nifA and ntrC genes of the stem nodulating bacterium ORS571, the nitrogen fixing symbiont of Sesbania rostrata: Regulation of nitrogen fixation (nif) genes in the free living versus symbiotic state

Summary A cosmid bank of ORS571, a diazotrophic bacterium capable of inducing aerial stem and root nodules on Sesbania rostrata, was constructed in the vector pLAFR1. A DNA probe carrying the Klebsiella pneumoniae nifA gene was used to identify nifA-and ntrC-like regions of ORS571 in the cosmid bank by colony hybridization. Cosmids carrying these regions were mapped by restriction endonuclease analysis, Southern blotting and transposon Tn5 mutagenesis. Selected Tn5 insertion mutations in the nifA/ntrC homologous regions were used for gene-replacement experiments and the resulting ORS571 mutants were examined for Nif, Fix and Ntr phenotypes. Two clearly distinct regulatory loci were thus identified and named nifA and ntrC. Plasmids carrying gene fusions of the ORS571 nifH and nifD genes to lacZ were constructed and the regulation of the ORS571 nifHDK promoter, and of the Rhizobium meliloti nifHDK promoter, was studied under varying physiological conditions in ORS571, ORS571 nifA::Tn5 and ORS571 nitrC::Tn5 strains. A model for the role of nifA and ntrC in the regulation of ORS571 nif and other nitrogen assimilation genes is proposed.     

Cloning and characterization of the pH 2.5 acid phosphatase gene, appA: Cyclic AMP mediated negative regulation

Summary The structural gene for an acid phosphatase coded for by the gene appA of Escherichia coli K12 was cloned from a cosmid library into pBR322 and the restriction map determined. Several appA deletion plasmids and a smaller appA ⁺ plasmid were constructed by in vitro recombination techniques and tested for their ability to complement an appA1 mutation. The appA gene was localized within a 2.1 kb segment. Its orientation was determined by construction of a hybrid plasmid carrying an appA-lacZ fusion. -galactosidase synthesized from the appA promoter was negatively regulated by cyclic AMP.     

The giant and complex genital plug of the asper group of Bothriurus (Scorpiones, Bothriuridae): morphology and comparison with other genital plugs in scorpions

The genital plugs of two species of the asper group of Bothriurus (Scorpiones: Bothriuridae) are described and compared with other genital plugs reported in the family Bothriuridae. In both species, B. asper and Bothriurus sp., the genital plug is cone-shaped and formed by fusion of the basal lobes of the hemispermatophore. Fusion is complete in B. asper and the surface of the plug has many microspines that anchor it to the female genital atrium. In Bothriurus sp., the basal lobes are partially fused, but free on the dorsal side, and the plug has a smooth surface with a dorsal curvature. Both genital plugs completely fill the genital atrium of inseminated females, pressing against the cuticular wall of the atrium. Given the large size and complex shape, the genital plug of the B. asper group is unique, not only among Bothriuridae, but in the order Scorpiones. This new type of genital plug resembles the genital plugs of the scorpion families Urodacidae and Vaejovidae. A comparison of the four major types of genital plugs reported in Bothriurus species and some other bothriurids is provided, as well as a comparison with other genital plugs reported in more distantly related families of scorpions.     

Plasmid pHH1 of Halobacterium salinarium: characterization of the replicon region, the gas vesicle gene cluster and insertion elements

The DNA sequence of the 5.7 kb plasmid pHH9 containing the replicon region of the 150 kb plasmid pHH1 from Halobacterium salinarium was determined. The minimal region necessary for stable plasmid maintenance lies within a 2.9 kb fragment, as defined by transformation experiments. The DNA sequence contained two open reading frames arranged in opposite orientations, separated by an unusually high AT-rich (60–70% A + T) sequence of 350 bp. All H. salinarium strains (H. halobium, H. cutirubrum) investigated harbour endogenous plasmids containing the pHH1 replicon; however, these pHH1-type plasmids differ by insertions and deletions. Adjacent to the replicon, and separated by a copy of each of the insertion elements ISH27 and ISH26, is the 9 kb p-vac region required for gas vesicle synthesis. Analysis of these and other ISH element copies in pHH1 revealed that most of them lack the target DNA duplication usually found with recently transposed ISH elements. These results underline the plasticity of plasmid pHH1.     

Bottom-up controls on a mixed-species HAB assemblage: A comparison of sympatric Chattonella subsalsa and Heterosigma akashiwo (Raphidophyceae) isolates from the Delaware Inland Bays, USA

Recent novel mixed blooms of several species of toxic raphidophytes have caused fish kills and raised health concerns in the highly eutrophic Inland Bays of Delaware, USA. The factors that control their growth and dominance are not clear, including how these multi-species HAB events can persist without competitive exclusion occurring. We compared and contrasted the relative environmental niches of sympatric Chattonella subsalsa and Heterosigma akashiwo isolates from the bays using classic Monod-type experiments. C. subsalsa grew over a temperature range from 10 to 30 °C and a salinity range of 5–30 psu, with optimal growth occurring from 20 to 30 °C and 15 to 25 psu. H. akashiwo had similar upper temperature and salinity tolerances but also lower limits, with growth occurring from 4 to 30 °C and 5 to 30 psu and optimal growth between 16 and 30 °C and 10 and 30 psu. These culture results were confirmed by field observations of bloom occurrences in the Inland Bays. Maximum nutrient-saturated growth rates (μ_max) for C. subsalsa were 0.6 d⁻¹ and half-saturation concentrations for growth (K_s) were 9 μM for nitrate, 1.5 μM for ammonium, and 0.8 μM for phosphate. μ_max of H. akashiwo (0.7 d⁻¹) was slightly higher than C. subsalsa, but K_s values were nearly an order of magnitude lower at 0.3 μM for nitrate, 0.3 μM for ammonium, and 0.2 μM for phosphate. H. akashiwo is able to grow on urea but C. subsalsa cannot, while both can use glutamic acid. Cell yield experiments at environmentally relevant levels suggested an apparent preference by C. subsalsa for ammonium as a nitrogen source, while H. akashiwo produced more biomass on nitrate. Light intensity affected both species similarly, with the same growth responses for each over a range from 100 to 600 μmol photons m⁻² s⁻¹. Factors not examined here may allow C. subsalsa to persist during multi-species blooms in the bays, despite being competitively inferior to H. akashiwo under most conditions of nutrient availability, temperature, and salinity.     

Foraging ecology and reproductive biology of the Stonechat Saxicola torquata: comparison between a revitalized, intensively cultivated and a historical, traditionally cultivated agro-ecosystem

An effective strategy to reintegrate biodiversity within otherwise intensively cultivated agroecosystems is to create set-aside and wildflower areas. It remains largely unknown, however, whether the ecological performance of an agroecosystem revitalized in this manner is comparable—from a species’ population biology perspective—to traditionally exploited farmland. To address this question we compared, during two successive years, the trophic ecology and breeding performance of an insectivorous, indicator passerine (the Stonechat Saxicola torquata) in a revitalized intensively cultivated farmland (RIC) and a traditional, extensively cultivated farmland (TEC) in southern Switzerland. The chicks' diet and prey abundance did not differ between the RIC and TEC, with orthopterans, caterpillars (Lepidoptera) and coleopterans predominating (approx. 80% of diet biomass). Although Stonechat pairs initiated more broods in TEC than in the RIC, reproductive success (number of fledglings/territory × year) did not differ significantly between the TEC and RIC. The chicks’ condition (body mass) was slightly better in TEC than in RIC, while no such effect could be shown for chick constitution (tarsus length) in either year. The inter-site (RIC vs. TEC) variation fell well within the inter-annual variation of breeding parameters, indicating that environmental stochasticity could be a greater determinant of reproductive output and young quality than agroecosystem type. Although in need of replication, these results suggest that incentives for setting aside farmland and creating wildflower areas within agroecosystems may not only enhance plant and invertebrate diversity, as has been demonstrated earlier, but can also support functioning populations of vertebrates situated at higher trophic levels along the food chain.