Transmission of Megatrypanum Trypanosomes to Cervus dama by Tabanidae1

Four fallow deer, Cervus dama, became infected with Trypanosoma (Megatrypanum) sp. by oral application of triturated guts from tabanids collected in an area with deer but without any cattle; four control calves remained negative. Upon challenge with triturated guts from tabanids from an area with pastured cattle, the four calves became infected with Trypanosoma (M.) theileri. The prepatent period in deer was five days or less. Haematopota spp. and Tabanus spp. were identified as vectors of the deer trypanosomes. It is concluded that the trypanosomes of C. dama belong to a Megatrypanum species that is not identical with T. theileri.     

Physical mapping of three fruit ripening genes：Endopolygalacturonase,ACC oxidase and ACC synthase from apple（Malus x domestica） in an apple rootstock A106（Malus sieboldii

The apple rootstock,A106(Malus sieboldii),had 17 bivalents in pollen mother cells at meiotic metaphase 1,and 17 chromosomes in a haploid pollen cell.Karyotypes were prepared from root-tip cells with 2n=34 chromosomes,Seven out of 82 karyotypes(8.5%) showed one pari of satellites at the end of the short arm of chromosome 3.C-bands were shown on 6 pairs of chromosomes 2,4,6,8,14,and 16 near the telomeric regions of short arms.Probes for three ripening-related genes from Malus x domestica:endopolygalacturonase(EPG,0.6kb),ACC oxidase(1.2kb),and ACC synthase(2kb)were hybridized in situ to metaphase chromosomes of A106.Hybridization sites for the EPG gene were observed on the long arm of chromosome 14 in 15 out of 16 replicate spreads and proximal to the centromere of chromosomes 6 and 11.For the ACC oxidase gene,hylridization sites were observed in the telomeric region of the short arm of chromosomes 5 and 11 in 87% and 81% of 16 spreads respectively,proxiaml to the centromere of chromosome 1 in 81% of the spreads,and on the long arm of chromosome 13 in 50% of the spreads. Physical mapping of three fruit ripening genes in an apple rootstock A106.Twenty five spreads were studied for the ACC synthase gene and hybridization sites were observed in the telomeric region of the short arm of chromosome 12 in 96% of the spreads.chromosomes 9 and 10 in 76% of the spreads,and chromosome 17 in 56% of the spreads.     

Partitioning and utilization of energy during the larval development of the xanthid crab, Rhithropanopeus harrisii (Gould)

An energy budget is constructed for the larval development of the crab Rhithropanopeus harrisii (Gould) fed nauplii of the brine shrimp Artemia salina (L.). Between the first zoeal instar and the megalopa, there is a 5.4-fold increase in caloric consumption and a 13.2-fold increase in dry weight. Weight specific energy content increases through the zoeal stages and drops at the megalopa. Rate of oxygen consumption increases steadily throughout development. Assimilation, gross growth, and net growth efficiencies increase steadily through zoeal development and drop at the megalopa. Calories put into tissue production exceed those expended via respiration in zoeal stages II–IV; the reverse is true in zoeal stage I and the megalopa.

A total energy budget has been calculated and the partitioning of energy is discussed in relation to other physiological studies on this species.     

The cytokineplast: purified, stable, and functional motile machinery from human blood polymorphonuclear leukocytes

We examined the formation of motile, chemotactically active, anucleate fragments from human blood polymorphonuclear leukocytes (PMN, granulocytes), induced by the brief application of heat. These granule-poor fragments are former protopods (leading fronts, lamellipodia) that become uncoupled from the main body of the cell and leave it, at first with a connecting filament that breaks and seals itself. The usual random orientation of such filaments can be controlled by preorientation of cells in a gradient of the chemotactic peptide, N-formylmethionylleucylphenylalanine (F-Met-Leu-Phe) (2x10(-9) M- 1x10(-8)). Cytochalsin B, 2.5-5 μg/ml, prevents fragment formation; colchicine, 10(-5) M, does not. In scanning electron micrographs, fragments are ruffled and the cell body rounded up and rather smooth. In transmission electron micrographs, fragments contain microfilaments but lack centrioles and microtubules. Like intact cells, both bound and free fragments can respond chemotactically to an erythrocyte destroyed by laser microirradiation (necrotaxis); the free, anucleate fragments may do so repeatedly, even after having been held overnight at ambient temperatures. We propse the name cytokineplast for the result of this self-purification of motile apparatus. The exodus of the motile machinery from the granulocyte requires anchoring of the bulk of the cell to glass and uncoupling, which may involve heat-induced dysfunction of the centrosome. In ultrastructural studies of the centrosomal region after heat, centriolar structure remains intact, but pericentriolar osmiophilic material appears condensed, and microtubules are sparse. These changes are found in all three blood cell types examined: PMN, eosinophil, and monocyte. Of these, the first two make fragments under our conditions; the more sluggish monocyte does not. Uncoupling is further linked to centrosomal dysfunction by the observation that colchicines-treated granulocytes (10(-5)M, to destroy the centrosome’s efferent arm) make fragments after less heat than controls. If motive force and orientation are specified mainly from the organelle-excluding leading front, then endoplasmic streaming in PMN is a catch-up phenomenon, and microtubules do not provide the vector of locomotion but rather stabilize and orient the “baggage” (nucleus, granuloplasm)—i.e., they prevent fishtailing. Moreover, constraints emanating from the centrosome may now be extended to include, maintenance of the motile machinery as an integral part of the cell.     

In vitro evaluation of native entomopathogenic fungi and neem (Azadiractha indica) extracts on Spodoptera frugiperda

The control of Spodoptera frugiperda is based on synthetic insecticides, so some alternatives are the use of entomopathogenic fungi (EF) and neem extract. The objective of the study was to evaluate in vitro effectiveness of native EF and neem extracts on S. frugiperda larvae. Six EF were identified by DNA sequencing of ITS regions from three EF (Fusarium solani, Metarrhizium robertsii, Nigrospora spherica and Penicillium citrinum). They were evaluated in concentrations of 1 × 10⁸ spores/ mL. In addition, a second bioassay was carried out evaluating only F. solani, M. robertsii and N. sphaerica and the addition of vegetable oil. On the other hand, extraction of secondary metabolites from neem seed (Azadirachta indica) was carried out by performing, mass (g) and solvent volume (mL ethanol and water) combinations, which were subjected to microwaves and ultrasound. Subsequently, these extracts were evaluated in concentrations of 3%, 4% and 5%. A survival analysis was performed for each of the bioassays. With respect to the results of the first bioassay, F. solani obtained a probability of survival of 0.476 on the seventh day, while in the second bioassay, M. robertsii obtained 0.488 survival probability. This suggests that the expected percentage of larvae that stay alive on the sixth day is 48.8%. However, in the evaluation of the neem extract the combination 1:12/70% to 4% caused 84% mortality of larvae. The use of native HE and neem extracts has potential for the control of S. frugiperda.     

Depth regulation of crab larvae in the absence of light

Behavioral responses to gravity and hydrostatic pressure have been investigated in two species of xanthid crabs Leptodius floridanus (Gibbes) and Panopeus herbstii Milne-Edwards to determine whether such responses provide a mechanism for depth regulation in the absence of light.In laboratory experiments, the four zoea stages and one megalopa stage of each species assume a differential vertical distribution in darkness, with succeeding stages showing a deeper overall distribution. Passive sinking rates increase in succeeding zoea stages and drop to an intermediate level after the molt to the megalopa stage. All zoea stages exhibit a negative geotaxis in the absence of light; the megalopa shows a positive geotaxis. The first zoea stage of Leptodius floridanus responds to an increase in hydrostatic pressure (up to 1 atmos above ambient) with an increase in swimming rate. This pressure response is shown to be reversible and not subject to short-term acclimation. The swimming rate of the last zoea stage does not increase in response to an increase in pressure.It is concluded that the responses of these larvae to gravity and hydrostatic pressure together with their characteristic passive sinking rates provide a mechanism for depth regulation in the absence of light that varies during ontogeny.     

The significance of diet in the growth and development of larvae of the blue crab,Callinectes sapidus rathbun,under laboratory conditions

The following protistan diets were tested on blue crab larvae: the algae Isochrisis galbana Parke, Monochrisis lutheri Droop, Dunaliella sp., and an unknown mixture; and the ciliated protozoans Euplotes vannus Muller and Parauronema virginianum(2/1) Thompson. None of these diets resulted in development past the first zoea stage, although some apparently were ingested and delayed mortality as compared to unfed controls.The rotifer Brachionus plicatilis Müller sustained good survival through early zoea development; however, rotifer-fed larvae did not metamorphose to the megalopa. Larvae of the polychaete Hydroides dianthus (Verrill) sustained crab larvae throughout zoea development, resulting in 17% survival to metamorphosis. The percentage mortality per stage was significantly lower in polychaete-fed larvae when compared with rotifer-fed larvae during zoea stages III, VI, and VII. Mean intermolt duration varied between diet treatments during the first three stages, but showed no differences during later zoea development. In tests on groups of late stage sibling larvae, Artemia salina L. nauplii gave development to metamorphosis, whereas rotifers did not.All the diets so far tested on blue crab larvae are classified according to their ability to sustain development. It is demonstrated that the two diets which allow completed development, Hydroides dianthus larvae and Artemia salina nauplii, contain 2–3 times as much lipid per dry weight as do rotifers. A metabolic requirement for lipid late in development may be indicated. Invertebrate larvae derived from yolky telolecithal and centrolecithal eggs may be an important dietary component for brachyuran larvae.     

Uptake of l-Phenylalanine in Synchronous Chlorella fusca. Characterization of the Uptake System

Phenylalanine uptake in Chlorella fusca was measured, using the membrane filter technique. The cells were synchronized, and harvested at specific points of the life cycle. Experiments with autospores showed that the uptake followed saturation kinetics, with a K_m= 5 μM. V_max, was 0.1 nmol/min × 10⁷ cells. The optimum temperature for the uptake was 40°C, and the activation energy was 1700 J/mol. The uptake showed a high specificity towards l -phenylalanine; presence of the unlabelled stereoisomer did not inhibit the uptake. Uptake of l -phenylalanine was inhibited in the presence of other analogues or other amino acids, but only if they were present in concentrations considerably higher than that of L-phenylalanine. Variations in the ratio of Na4⁺ to K⁺ in the external solution during uptake experiments did not have any influence upon the uptake rate of l -phenylalanine. The cells were able to take up the amino acid against a concentration gradient. At pool maximum the ratio between internal and external amino acid concentration was 1000/1. 2,4-Dinitro-phenol inhibited the uptake completely. Exchange between internal and external l -phenylalanine could not be demonstrated. The K_m value did not change during the life cycle of the cells. The uptake rate reached a maximum at the end of the light period, and fell to a minimum just before sporulation started. It is concluded that Chlorella fusca cells have a highly specific, active uptake system for l -phenylalanine. The system is constitutive, independent on the K or Na concentration, and the mechanism of uptake does not change during the life cycle of the cells.     

HISTOCHEMICAL STUDIES ON MUCOPROTEINS IN NERVE CELLS OF THE DOG

Autonomic and sensory ganglion cells in the senile dog contain a deposition of PAS-positive substances which has been shown to be mucoprotein in nature. Data are presented to show that this PAS-positive mucoprotein can be demonstrated by metachromatic staining with toluidine blue after the mucoprotein is sulfated. This procedure indicates that mucoprotein is also present in a granular form in all nerve cells in both senile and young dogs. The evidence for this is further substantiated by the use of the aldehyde-fuchsin stain following both periodic acid oxidation and sulfation. The granular and non-granular deposition can be demonstrated by the periodic acid-aldehyde-fuchsin method due to the affinity of the aldehyde-fuchsin stain for aldehydes. It can be demonstrated following the sulfation-aldehyde-fuchsin method owing to the affinity of the stain for the sulfuric group. The evidence for this latter phenomenon has been reported by Scott and Clayton (6). It is concluded that mucoprotein is present in a granular form in all nerve cells in both senile and young dogs but is not concentrated enough in the latter to be demonstrated by the PAS method.