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Importance of algal biomass to growth and development of Anopheles gambiae larvae.

We conducted experiments to investigate the importance of algal food resources for larval growth and adult emergence of Anopheles gambiae Giles s.s. in simulated larval habitats in Kenya, and in greenhouse and laboratory microcosms in the United States. In the first experiment, we used shading to reduce algal biomass, and because algal production and larval development might be a function of underlying soil nutrients, we crossed sun-shade treatments with soils of two distinct types collected near larval habitats. Shading reduced pupation rates and total adult biomass of An. gambiae by approximately 50%. Soil type had no significant effect on mosquito production, but it did significantly affect concentrations of phosphorus and chlorophyll a in the surface microlayer. In a subsequent experiment conducted in the greenhouse to reduce temperature differences found between the shaded and sunlit treatments, <1% of larvae in the shaded treatments reached the pupal stage. There was a marked reduction of chlorophyll a levels as a function of shading and larval density. In a third experiment, larvae receiving material harvested from sunlit surface microlayers performed as well as those receiving liver powder, whereas those receiving surface microlayer from shaded habitats suffered >90% mortality and failed to pupate. In a fourth experiment, glucose was added to shaded microcosms to stimulate bacterial activity in the absence of algae. Bacterial growth rates were 2 to 3 times higher, and larval development was enhanced in glucose-amended treatments. However, pupation rates and adult weights in glucose-amended shaded microcosms were still poor compared with those in nonamended sunlit microcosms. Overall, these results demonstrate the importance of algal biomass in the surface microlayers of larval habitats to development and adult production of An. gambiae.

Animal Feed↗

Effect of two thermal regimes on the muscle growth dynamics of sea bass larvae, Dicentrarchus labrax L.

Muscle growth was studied in larvae of sea bass, Dicentrarchus labrax L., reared at two temperatures: real ambient temperature ( congruent with 15 degrees C during vitelline phase and increased gradually) and 19 degrees C from fertilization until the end of larval development. Muscle cellularity, body length and body weight were measured. Early temperature influenced larval development and so, pre-larval phase finished earlier at 19 degrees C than at ambient temperature (4 and 6 days, respectively). Temperature also affected muscle growth such that at hatching and at mouth opening hypertrophy of muscle fibres was greater at 19 degrees C (P < 0.05), whereas hyperplasia was similar in both groups. After 25 days, the cross-sectional area of the white muscle was greater at 19 degrees C (P < 0.05), which was mainly associated with a higher proliferation of new white muscle fibres. At this stage the body length was also higher at 19 degrees C. Metamorphosis finished earlier in fish reared at 19 degrees C (52 days) than at natural temperature (82 days). At this developmental stage body length and cross-sectional area of the myotome were similar in both groups. However, muscle cellularity differed between groups. Thus, hypertrophy of muscle fibres was higher in fish reared at ambient temperature (P < 0.05), whereas proliferation of new muscle fibres was higher at 19 degrees C (P > 0.05).

Animals↗

Photoperiodism and control of summer diapause in the Mediterranean tiger moth Cymbalophora pudica.

Photoperiodic responses to both constant and changing photoperiods were studied in the Mediterranean tiger moth Cymbalophora pudica. Embryos, larval instars and prepupae were all sensitive to photoperiod, and the responses of larvae and prepupae to changing photoperiods were similar. At 23+/-2 degrees C, constant 24-h photoperiods with short photophases (11, 12h) induced a long diapause (mean 88days) whereas long photophases (14, 16h) induced a short diapause (mean 52days). A change to a longer or shorter photophase during larval development or during diapause caused a significant extension (up to a maximum of 138days) or shortening (down to a minimum of 10days) of diapause, respectively, but only when at least one of the photophases was longer than 14h. Thus, shortening and prolongation of photophase had an opposite effect than constant short and long photophases, respectively. Changes within the range of photophases of 10-14h did not cause a significant change in diapause duration.Experimental results enabled us to outline the mechanisms regulating voltinism and the duration of summer diapause. For the monovoltine cycle, cold autumn/winter temperatures slow down larval development, and prepupal aestivation starts relatively late (March, April). Prepupae are then kept in diapause by the increasing daylength (>14h after late April). Pupation is synchronized by decreasing daylength after summer solstice, and imagoes emerge in September/October. For the bivoltine cycle, when the autumn/winter temperatures are relatively warm, a certain portion of the population (depending on the individual rate of growth) may be diverted to a bivoltine life-cycle. In such a case, larval development is fast and short enough to allow an early start of diapause (prior to or during February). The duration of such early diapause is not influenced by changes in daylength (<14h); pupation occurs very early (April/May), and spring generation imagoes fly and oviposit in May/June. Summer larvae and prepupae live under decreasing daylength (>14h), which shortens their diapause to 20-30days. Imagoes of the autumnal generation thus occur in September/October, together with the univoltine portion of the population.

Journal Article↗

The embryonic development of larval muscles in Drosophila.

Each of the abdominal hemisegments A2-A7 in the Drosophila larva has a stereotyped pattern of 30 muscles. The pattern is complete by 13 h after egg laying, but the development of individual muscles has begun with the definition of precursors at least by the onset of germ band shortening, some 5.5 h earlier. The earliest signs of muscle differentiation are cell fusions, which occur in the ventralmost mesoderm overlying the CNS and at stereotyped positions in the rest of the mesoderm as the germ band shortens. At the end of shortening, the pattern of muscle precursors produced by these fusions is complete. Precursors filled with dye reveal extensive fine processes probably involved initially in cell fusion and, subsequently, in navigation over the epidermis to form attachment points. The muscle pattern is formed before innervation and without cell death. Thus, neither of these processes is involved in determining the distribution of precursors. Evidence is presented for the view that the development of the larval muscle pattern in Drosophila depends on a prior segregation of founder cells at appropriate locations in the mesoderm with which other cells fuse to form the precursors.

Animals↗

Development of Drosophila larval neuromuscular junctions: maintaining synaptic strength.

In spite of the available information about the development of Drosophila neuromuscular junctions, the correlation between nerve terminal morphology and maintenance of synaptic strength has still not been systematically addressed throughout larval development. We characterized the growth of the abdominal longitudinal muscle 6 (m6) and the motor terminals Ib and Is that innervate it within segment 4. In addition, we measured the evoked excitatory junction potential (EJP) amplitudes while the Ib and Is axons were selectively recruited. Regression analysis with natural log transformation of response variables indicated that the developmental curves for m6 and the motor axons Ib and Is were best fitted as second order polynomial regressions during larval development. Initially Is terminals are longer and possess more synaptic varicosities at the first instar stage. The Is terminals also grow faster in subsequent developmental stages. The growth of nerve terminals and their target m6 are not proportional although tightly correlated. This results in a larger average muscle area innervated by a single varicosity as the animal develops. The amplitudes of the EJPs of Ib and Is neurons show no developmental difference in their amplitudes from the first to the late third larval instar. The Is axon consistently produced larger EJPs than the Ib axon at each developmental stage. The time constants for both rising and decay phases of EJPs increase exponentially throughout larval development. The results presented not only help in quantifying the normal development of Drosophila neuromuscular junctions, but also provide a framework for future investigations to properly interpret developmental abnormalities that may occur in various mutants.

Animals↗

[RNA/DNA ratio as an index of physiological condition of Colossoma macropomum and Piaractus branchypomus (Pisces: Characiformes) during embryonic development].

We evaluated RNA/DNA ratio as an index of physiological condition during larval development of a hybrid between the fishes Colossoma macropomum (cachama) and Piaractus brachypomus (morocoto). The samples were obtained by induced reproductive technology and the eggs were maintained in acrylic conical incubator with a continuous waterflow. Embryonic development, from egg fertilization to cell division and hatch out, took 12 hours 20 minutes at 29.5 degrees C, dissolved oxygen contents of 6.0 ppm and pH 7.5. Nucleic acids quantification was determined by fluorometry with ethidium bromide and Hoechst 33258 dyes. We observed significant changes of RNA/DNA ratios during all stages of the embryonic larval development. Therefore, RNA/DNA relation is an useful technique to evaluate physiological condition in short period and could be utilized as nutritional condition and/or instantaneous growth for routine check to verify the health status in early life of cultivated species.

Animals↗

Correlated responses to selection on body size in Drosophila melanogaster.

Correlated responses to artificial selection on body size in Drosophila melanogaster were investigated, to determine how the changes in size were produced during development. Selection for increased thorax length was associated with an increase in larval development time, an extended growth period, no change in growth rate, and an increased critical larval weight for pupariation. Selection for reduced thorax length was associated with reduced growth rate, no change in duration of larval development and a reduced critical larval weight for pupariation. In both lines selected for thorax length and lines selected for wing area, total body size changed in the same direction as the artificially selected trait. In large selection lines of both types, the increase in size was achieved almost entirely by an increase in cell number, while in the small lines the decrease in size was achieved predominantly by reduced cell size, and also by a reduction in cell number. The implications of the results for evolutionary-genetic change in body size in nature are discussed.

Animals↗

Transovarial transmission efficiency of Babesia bovis tick stages acquired by Rhipicephalus (Boophilus) microplus during acute infection.

The protozoan parasite Babesia bovis, a reemerging threat to U.S. cattle, is acquired by adult female ticks of the subgenus Boophilus and is transovarially transmitted as the kinete stage to developing larval offspring. Sporozoites develop within larvae and are transmitted during larval feeding on a bovine host. This study evaluated the efficiency of B. bovis infection within Rhipicephalus (Boophilus) microplus following acquisition feeding on acutely parasitemic cattle. Parasite levels were quantified in blood from experimentally infected cattle and within hemolymph and larvae derived from acquisition-fed female B. microplus. There was a positive correlation between blood parasite levels in acutely parasitemic cattle and kinete levels in the hemolymph of adult female Boophilus ticks following acquisition feeding; however, there was no relationship between kinete levels in females and infection rates of larval progeny. Boophilus microplus females that acquisition fed produced larval progeny with infection rates of 12% to 48%. Importantly, larvae derived from replete females with very low levels of kinete infection, as demonstrated by microscopy and PCR, had infection rates of 22% to 30% and transmitted B. bovis during transmission feeding. These data demonstrate that although hemolymph infection may be undetectable, transmission to larval progeny occurs at a level which ensures transmission to the bovine host.

Acute Disease↗

The effect of grazing forage containing condensed tannins on gastro-intestinal parasite infection and milk composition in Angora does.

The objective of this study was to evaluate effects of the condensed tannin (CT)-containing forage sericea lespedeza (sericea lespedeza (SL); Lespedeza cuneata; 15.2% CT), on fecal egg count (FEC), larval development (larvae/10 g of feces), worm burden and immune response compared with a crabgrass (Digitaria ischaemum)/Kentucky 31 tall fescue (Festuca arundinacea; control forage (CTF)) forage low in CT (0.32% CT) in grazing Angora does and their kids. Fifty worm-free mixed-age does were randomly allocated to three treatments. One treatment (10 does; initial liveweight (LW) = 45+/-1.5 kg) entailed grazing of SL forage from April 25 to July 15, 2002 with a second treatment of CTF (20 does; initial LW = 43+/-1.4 kg) grazing during the same period. Does of the third treatment (20 does; initial LW = 44+/-1.4 kg) grazed a sward of SL for 2 weeks and then one of CTF for 2 weeks followed by alternating between the two pastures every 2-week rotational grazing (ROT). To gauge levels of infective larvae on pasture, three worm-free Angora kids (initial LW = 3.6+/-0.2 kg) were randomly selected as tracers. Tracers grazed for final 60 days and were euthanized for determination of worm burden. The immune response of does was measured by skin thickness reaction after the intradermal injection of 250 microg phytohemagglutinin (PHA). Mean FEC for SL and ROT were substantially lower (P < 0.01) than for CTF does (145, 329 and 894 eggs/g, respectively). The FEC for kids was lower (P < 0.05) for SL than for ROT and CTF (550, 2757 and 3600 eggs/g, respectively). Total fecal egg output (3.3, 6.0 and 26.9 x 10(5) eggs/day, respectively) and larval development (242, 263 and 792 larvae/10 g, respectively) were lower (P < 0.05) for SL and ROT than for CTF. Tracers grazing on SL had lower total worm burdens than ROT and CTF (P < 0.01). The immune response was higher (P < 0.01) for SL (4.9 mm) and ROT (6.0 mm) than for CTF (3.0 mm) at 12 h after injection of PHA. The packed cell volume (PCV) in does was higher (P < 0.01) for SL and ROT than for CTF (27, 26 and 23%, respectively). Does that grazed CT-containing forage had considerably lower milk somatic cell counts (SCC) than does grazing non-CT-containing forage. In summary, grazing CT forages reduced FEC, larval development and worm burden, and also appeared to enhance immune response. The CT-containing forage SL reduced gastro-intestinal parasite infections of Angora does and kids.

Animal Feed↗

Are increased nutrient inputs responsible for more outbreaks of crown-of-thorns starfish? An appraisal of the evidence.

The cause(s) of primary outbreaks of the coral-eating crown-of-thorns starfish (Acanthaster planci) are still subject to scientific controversy. The possibility of primary outbreaks being linked to terrestrial runoff has been postulated a number of times, suggesting that enhanced nutrient supply is critical for enhanced A. planci larval development. This paper examines the evidence for such a cause, focussing particularly on the Great Barrier Reef (GBR). Nutrient discharges from rivers have increased at least four-fold in the central GBR over the last century, and concentrations of large phyto-plankton (>2 microm) of the inshore central GBR shelf in the wet season when A. planci larvae develop, is double that of other places and times. Larval development, growth and survival increase almost ten-fold with doubled concentrations of large phyto-plankton. This and other lines of evidence suggest that frequent A. planci outbreaks on the GBR may indeed be a result of increased nutrient delivery from the land.

Animals↗

Effects of prolactin and bromocriptine in Discoglossus pictus (anuran amphibian. Otth) tadpoles.

1. The results obtained by ovine prolactin administration during the larval development of Discoglossus pictus (OTT) suggests that prolactin-like hormone is the "larval factor" necessary for growth and for maintaining the larval aquatic features. 2. Bromocriptine treatment during the larval development of D. pictus has contrary effect on growth and larval structures. 3. Prolactin administration does not inhibit but delays metamorphosis. 4. When bromocriptine is added at late stages of the development, metamorphosis is accelerated.

Animals↗

Epidermal cell cycle and region-specific growth during segment development in Artemia.

During larval development in instar IV brine shrimp, segment 1 grew by cell replication and cell differentiation. Cell cycle analysis revealed that the cell cycle was synchronized with the molt cycle. Mitosis occurred late in the instar and S phase began at hr 6 of the following instar. Three populations of cells comprised the dorsal integument. The medial-dorsal region did not grow. Cell enlargement occurred in the dorsal-lateral population while cell replication took place in the lateral population. The limb bud (ventral surface) grew in width by replication in the distal population, and in height by enlargement of the general epidermal cells in the proximal population. Expansion of each region of the integument was proportional to the cell growth in that region. Moreover, both growth processes were dependent on the level of nutrients and were enhanced by diets enriched in polyunsaturated fatty acids. A nutrient-dependent growth control point occurred in the G1 period. The commitment to replication and differentiation occurred by hr 2 of instar IV. The findings show that integumental growth is a result of region-specific cell growth processes which are controlled by nutrients during the G1 period.

Animal Nutritional Physiological Phenomena↗

Relationships among salinity, egg size, embryonic development, and larval biomass in the estuarine crab Chasmagnathus granulata Dana, 1851.

We studied interrelationships between initial egg size and biomass, duration of embryogenesis at different salinities, and initial larval biomass in an estuarine crab, Chasmagnathus granulata. Ovigerous females were maintained at three different salinities (15 per thousand, 20 per thousand and 32 per thousand); initial egg size (mean diameter), biomass (dry weight, carbon and nitrogen) as well as changes in egg size, embryonic development duration, and initial larval biomass were measured.Initial egg size varied significantly among broods from different females maintained under identical environmental conditions. Eggs from females maintained at 15 per thousand had on average higher biomass and larger diameter. We hypothesise that this is a plastic response to salinity, which may have an adaptive value, i.e. it may increase the survivorship during postembryonic development. The degree of change in egg diameter during the embryonic development depended on salinity: eggs in a late developmental stage were at 15 per thousand significantly larger and had smaller increment than those incubated at higher salinities. Development duration was longer at 15 per thousand, but this was significant only for the intermediate embryonic stages. Initial larval biomass depended on initial egg size and on biomass loss during embryogenesis. Larvae with high initial biomass originated either from those eggs that had, already from egg laying, a high initial biomass (reflecting individual variability under identical conditions), or from those developing at a high salinity (32 per thousand), where embryonic biomass losses were generally minimum. Our results show that both individual variability in the provisioning of eggs with yolk and the salinity prevailing during the embryonic development are important factors causing variability in the initial larval biomass of C. granulata, and thus, in early larval survival and growth.

Journal Article↗

Interactions of temperature and steroids on larval growth, development, and metamorphosis in a toad (Bufo boreas).

The effects of temperature and steroids [testosterone (T), estradiol (E2), and corticosterone (B)] on premetamorphic growth and development were investigated in the toad (Bufo boreas). The effects of steroids were both temperature and age dependent. In the first experiment, steroids (1.1-1.4 microM) were administered by dissolving them in the water beginning 1 day after hatching at 22 degrees C or 27 degrees C. At 22 degrees C, B inhibited growth (P < 0.001) but had no significant effect on development. Forelegs never emerged in B-treated animals and all died before complete tail resorption. Discontinuation of B treatment allowed normal growth and metamorphosis, but the resulting post-metamorphic animals were significantly shorter (snout-vent length, P < 0.001) than after other treatments. At 22 degrees C, T and E2 had no effect on larval growth and development or size at metamorphosis (P > 0.05), but T induced early foreleg emergence (FLE) (P < 0.005). At 27 degrees C, B was fatal after 2 weeks of treatment, and T and E2 inhibited growth (P < 0.001) and development (P < 0.001), but did not affect time to FLE. In a second experiment at 27 degrees C, treatment with 1.1 microM B starting 15 days after hatching induced early metamorphic events (P < 0.001), such as tail resorption and emergence of the left foreleg (but not the right), but jaw and head restructuring failed to occur. All B-treated animals died before complete tail resorption. In a third experiment, 0.275 and 1.11 microM B, starting at day 43 (stage 43), induced early FLE (P < 0.05) and decreased snout-vent length at tail resorption (P < 0.005) without a dose effect. A higher dose of B (4.44 microM) decreased snout-vent length at tail resorption and time to FLE (P < 0.05) but did not affect body weight at metamorphosis (P > 0.05). Animals in this experiment survived to complete tail resorption and transformed normally. The actions of B in these experiments closely resemble those observed with administration of thyroid hormones, suggesting that steroids may interact with endogenous thyroid hormones.

Animals↗

Trade-offs during the development of primary and secondary sexual traits in a horned beetle.

Resource allocation trade-offs during development affect the final sizes of adult structures and have the potential to constrain the types and magnitude of evolutionary change that developmental processes can accommodate. Such trade-offs can arise when two or more body parts compete for a limited pool of resources to sustain their growth and differentiation. Recent studies on several holometabolous insects suggest that resource allocation trade-offs may be most pronounced in tissues that grow physically close to each other. Here we examine the nature and magnitude of developmental trade-offs between two very distant body parts: head horns and genitalia of males of the horned scarab beetle Onthophagus taurus. Both structures develop from imaginal disklike tissues that undergo explosive growth during late larval development but differ in exactly when they initiate their growth. We experimentally ablated the precursor cells that normally give rise to male genitalia at several time points during late larval development and examined the degree of horn development in these males compared to that of untreated and sham-operated control males. We found that experimental males developed disproportionately larger horns. Horn overexpression was weakest in response to early ablation and most pronounced in males whose genital disks were ablated just before larvae entered the prepupal stage. Our results suggest that even distant body parts may rely on a common resource pool to sustain their growth and that the relative timing of growth may play an important role in determining whether, and how severely, growing organs will affect each other during development. We use our findings to discuss the physiological causes and evolutionary consequences of resource allocation trade-offs.

Animals↗

Neuronal development in larval chiton Ischnochiton hakodadensis (Mollusca: Polyplacophora).

Chitons are the most primitive molluscs and, thus, a matter of considerable interest for understanding both basic principles of molluscan neurogenesis and phylogeny. The development of the nervous system in trochophores of the chiton Ischnochiton hakodadensis from hatching to metamorphosis is described in detail by using confocal laser scanning microscopy and antibodies raised against serotonin, FMRFamide, and acetylated alpha tubulin. The earliest nervous elements detected were peripheral neurons located in the frontal hemisphere of posthatching trochophores and projecting into the apical organ. Among them, two pairs of unique large lateral cells appear to pioneer the pathways of developing adult nervous system. Chitons possess an apical organ that contains the largest number of neurons among all molluscan larvae investigated so far. Besides, many pretrochal neurons are situated outside the apical organ. The prototroch is not innervated by larval neurons. The first neurons of the developing adult central nervous system (CNS) appear later in the cerebral ganglion and pedal cords. None of the neurons of the larval nervous system are retained in the adult CNS. They cease to express their transmitter content and disintegrate after settlement. Although the adult CNS of chitons resembles that of polychaetes, their general scenario of neuronal development resembles that of advanced molluscs and differs from annelids. Thus, our data demonstrate the conservative pattern of molluscan neurogenesis and suggest independent origin of molluscan and annelid trochophores.

Animals↗

The wings of Bombyx mori develop from larval discs exhibiting an early differentiated state: a preliminary report.

Lepidopteran insects present a complex organization of appendages which develop by various mechanisms. In the mulberry silkworm, Bombyx mori a pair of meso- and meta-thoracic discs located on either side in the larvae gives rise to the corresponding fore- and hind-wings of the adult. These discs do not experience massive cell rearrangements during metamorphosis and display the adult wing vein pattern. We have analysed wing development in B. mori by two approaches, viz., expression of patterning genes in larval wing discs, and regulatory capacities of larval discs following explantation or perturbation. Expression of Nubbin is seen all over the presumptive wing blade domains unlike in Drosophila, where it is confined to the hinge and the wing pouch. Excision of meso- and meta-thoracic discs during the larval stages resulted in emergence of adult moths lacking the corresponding wings without any loss of thoracic tissues suggesting independent origin of wing and thoracic primordia. The expression of wingless and distal-less along the dorsal/ventral margin in wing discs correlated well with their expression profile in adult Drosophila wings. Partially excised wing discs did not show in situ regeneration or duplication suggesting their early differentiation. The presence of adult wing vein patterns discernible in larval wing discs and the patterns of marker gene expression as well as the inability of these discs to regulate growth suggested that wing differentiation is achieved early in B. mori. The timings of morphogenetic events are different and the wing discs behave like presumptive wing buds opening out as wing blades in B. mori unlike evagination of only the pouch region as wing blades seen in Drosophila.

Animals↗

The biology of Ctenocephalides canis in Ireland.

A colony of Ctenocephalides canis was established using dogs as hosts. Two diets were used as media. Fleas reared on cats did not develop beyond the first larval stages. The effects of different temperatures on egg hatching and larval development were examined. Larval survival was poor at 22 degrees C and 25 degrees C at 50% relative humidity, but good at 75% relative humidity at these temperatures. The development from egg to adult took 21 days.

Animals↗