Bemerkungen zur Biologie sowie Haltung und Vermehrung des Sokotra-Riesengeckos, Haemodracon riebeckii (Peters, 1882)

The Socotra Giant Gecko, Haemodracon riebeckii, is the largest species of lizard on Socotra Island. The nocturnal, arboreal and rupiculous living geckos are omnivorous. Two pairs were kept in terrariums and were fed with various insects (crickets, locusts, cockroaches), sweet fruits and other feeding stuff (such as meat, fish). Temporarily H. riebeckii was kept together with other lizards (Eublepharis macularius, Trachylepis socotrana), without any signs of aggressive behaviour. Juveniles and adults of both sexes are able to produce a sound. These acoustic signals seem to be related to predators, because never any intraspecific function could be observed. Within seven years of captive breeding two females produced 253 eggs. Usually two white and sticky soft-shelled eggs were laid within one clutch, more rarely a single egg was laid. The two eggs of a clutch were always laid on the same day. H. riebeckii belongs to the geckos that bury their eggs and practice some brood care, but no special parental care. The female is able to proof with her hind legs the deep and shape of a hollow in the substrate to bury the eggs, which were buried in a sticky and soft-shelled condition. They are oval in shape (egg length 16.4-19.8 mm, egg width 12.4-17.8 mm, quotient EL:EW 1.22±0.05) and have in the beginning a weight ranging from 1.7100 to 2.5201 g. As typical for geckos with hard-shelled eggs the egg weight decreases during the incubation period. The loss can be between 5.59 to 30.29%. The development of eggs up to hatching of young depends upon temperature and the germinal stage in the laid egg. The time difference between the hatching of the young within one clutch of two eggs was usually 1 to 5 days. In some cases there were, however, longer differences (up to 61 days), which are probably caused by different developmental stages of the embryos during the time of egg laying. The shortest incubation period recorded during our investigations was 83 days for eggs incubated at constant temperature of 28 to 29.5 °C and the longest 359 days at 26 to 26.5 °C. Constant high incubation temperatures caused a premature hatching of young. In normal hatched young were the yolk sac retracted and the navel closed. In premature hatched young were the yolk not resorbed and the mortality within the first three month comparatively high. The snout-vent length (SVL) of newly hatched young is from 27 to 39 mm and the tail length (TL) from 25 to 38 mm (SVL:TL index 0.90-1.27), the weight is from 0.7688 to 1.5366 g. Young specimens are distinguished from adults by the brown/white striped lower jaw and the white-banded tail. Young which hatched in the terrarium were eaten by the adults. A loss of young can be avoided if they are raised individually.     

Near anoxia and sulfide as possible factors influencing the spatial distribution of Acartia tonsa and Acartia clausi: Comparative evaluation of egg tolerance

In many coastal and estuarine areas the planktonic copepods Acartia tonsa and Acartia clausi show a spatial separation with A. tonsa restricted to brackish waters and confined environments and A. clausi inhabiting areas more influenced by sea water. The hatching and viability of A. tonsa and A. clausi eggs exposed to anoxia and anoxia/sulfide (conditions that are frequent in bottom waters of the most confined areas) was evaluated to determine if these stress factors play a role in the distribution of these species. Subitaneous eggs, spawned by laboratory reared organisms, were incubated in near anoxia (< 7.59 × 10^− 3 mmol O₂ L^− 1) or anoxia/sulfide (∼ 1 mmol L^− 1) for different periods (1, 4, 8 and 15 days), then transferred to normoxic conditions. The exposure of the eggs to near anoxia and sulfide appears to induce the same response (quiescence) in both species. Exposure times ≤ 8 days to near anoxia or anoxia/sulfide did not affect egg viability, while 15 day exposure caused significant declines in hatching success of both species. A significant difference between the effects of near anoxia and anoxia/sulfide was observed when incubation lasted 15 days; hatching of eggs exposed to sulfide being higher than that of eggs exposed to near anoxia for both species. No significant differences were observed between the two species in hatching success of eggs exposed to both near anoxia and anoxia/sulfide (with the exception of eggs incubated in near anoxia for 4 days). The results indicate that the impact of anoxia and sulfide on the eggs of the two Acartia species cannot be a factor explaining the spatial distribution in coastal and brackish environments of these copepods.Feeding experiments on A. clausi were also performed. Suitability of different algal species to rear this copepod was evaluated and the results were compared with data previously obtained for A. tonsa. Differences in feeding needs between A. clausi and A. tonsa are discussed.     

Metallothionein and cellular energy allocation in the estuarine mysid shrimp Neomysis integer exposed to cadmium at different salinities

In the present study the induction of metallothioneins (MTs) and effects on the cellular energy allocation (CEA) in euryhaline crustacean Neomysis integer exposed to Cd at different salinities were studied. N. integer was exposed to the same sub-lethal concentration of free cadmium ion (1/5 of the cadmium activity of the reported 96 h LC₅₀ value) in hypo-osmotic (7.2 μg Cd/L at 5 psu), isosmotic (23.0 μg Cd/L at 16 psu) and hyper-osmotic media (38.1 μg Cd/L at 25 psu) for 7 days. By using the free Cd concentration as the basis for conducting the exposures, the effect of salinity on cadmium speciation was eliminated and therefore the true effect of salinity as an abiotic factor on the MT induction and CEA could be studied.MT content was quantified by differential pulse voltammetry (DPV); this is the first time that this method is applied to assays with N. integer. No significant differences in MT levels between the control and Cd-exposed groups were observed. The measured MT levels (ranging from 32.3 to 75.7 μg/mg_{(cytosolic protein)}) probably represent the constitutive MT levels responsible for binding essential metals. The differences in MT levels observed at the different salinities indicate a possible relationship between some physiological process (possibly osmoregulation) other than detoxification and MT induction. A decrease in salinity caused an increase in MT level in N. integer, and this was significantly correlated to the CEA values (Spearman correlation coefficient r = − 0.56, p = 0.016). Our results indicate that salinity can change the energy status and MT content of N. integer. These findings should be taken into account when using these biomarkers - and especially MTs - in field studies and environmental monitoring.     

Fasciola gigantica: Cercarial shedding pattern from Lymnaea natalensis after long-term exposure to cadmium at different temperatures

Little is known about the response of Fasciola spp., inside its snail host to pollutant toxicity. The effect of long-term exposure of Lymnaea natalensis to 0.1 μg/l, 10 μg/l, or 100 μg/l cadmium (Cd) on the infection rates with F. gigantica, percentage of cercariae-shedding snails and cercariae shedding pattern was investigated. The snails were exposed to Cd for 7 days either pre-infection with single Fasciola miracidium or during the late pre-patent period of the infection. The possible interaction between metal exposure and acclimatization temperature was also studied in three ranges; 16-18 °C, 23-25 °C and 30-32 °C. Results clearly showed that host exposure to 100 μg/l Cd significantly affected the pattern of Fasciola development inside its snail host, and that the acclimatization temperature was a key factor affecting the role played by Cd. Pre-infection exposure to Cd caused a significant reduction in the infection rates where the effect was temperature-dependent. Post-infection exposure to Cd significantly increased the percentage of cercariae-shedding snails; the effect was temperature-independent. Interestingly, post-infection exposure significantly altered the differential cercarial output pattern, while no clear impact on the total was found. The alteration in the differential cercarial output was represented as a significantly higher proportion of floating metacercaria as compared to fixed ones and also higher proportion of dead cercariae which may directly reduce the transmission of Fasciola to the final host. The mechanisms of cadmium impact are briefly discussed.     

Modulation of ethanol toxicity by Asian ginseng (Panax ginseng) in Japanese ricefish (Oryzias latipes) embryogenesis

Alcohol consumption by women during pregnancy often induces fetal alcohol spectrum disorder (FASD) in children who have serious central nervous system (CNS), cardiovascular, and craniofacial defects. Prevention of FASD, other than women abstaining from alcohol drinking during pregnancy, is not known. A limitation of the use of synthetic anti-alcoholic drugs during pregnancy led us to investigate herbal products. In particular, many plants including Asian ginseng (Panax ginseng) have therapeutic potential for the treatment of alcoholism. We used Japanese ricefish (medaka) (Oryzias latipes), an animal model of FASD, for identifying herbal medicines that can attenuate ethanol toxicity. Fertilized eggs in standard laboratory conditions were exposed to ginseng (PG) root extract (0–2 mg/mL) either 0–2 (group A) or 1–3 (group B) day post fertilization (dpf) followed by maintenance in a clean hatching solution. The calculated IC₅₀ as determined 10 dpf in A and B groups were 355.3 ± 1.12 and 679.7 ± 1.6 μg/mL, respectively. Simultaneous exposure of embryos in sub-lethal concentrations of PG (50–200 μg/mL) and ethanol (300 mM) for 48 h disrupted vessel circulation and enhanced mortality. However, PG (100 μg/mL) may partially protect trabecular cartilage (TC) deformities in the neurocranium in B group embryos induced by ethanol (300 mM). To understand the mechanism, embryonic ethanol concentration was measured at 2 dpf and adh5, adh8, aldh2, aldh9a, catalase, GST, and GR mRNAs were analyzed at 6 dpf. It was observed that although ethanol is able to reduce adh8 and GST mRNA contents, the simultaneous addition of PG was unable to alter ethanol level as well as mRNA contents in these embryos. Therefore, antagonistic effects of PG on ethanol toxicity are mediated by a mechanism which is different from those regulating ethanol metabolism and oxidative stress.     

Effect of pulsed exposure to heavy metals (copper and cadmium) on some population variables of <Emphasis Type="Italic">Brachionus calyciflorus</Emphasis> Pallas (Rotifera: Brachionidae: Monogononta)

Heavy metals are widely recognized as potential toxic agents to zooplankton, yet experiments are usually performed with a continuous exposure to the metal being analyzed. Here we describe experiments that examined the influence of pulsed exposure of the heavy metals copper and cadmium to a parthenogenetic population of the planktonic rotifer Brachionus calyciflorus. Our protocol called for exposure durations of 3, 6, 12, and 24 h to either copper (as CuSO₄) at concentrations of 0.0375, 0.075, 0.15 mg l⁻¹ or cadmium (as CdCl₂) at concentration of 0.025, 0.05, 0.1 mg l⁻¹. Control animals were treated in similar ways but did not receive exposure to heavy metals. Four end points were used to evaluate the outcome of exposure: population growth (r), body size, egg ratio, and egg hatching percent. Increase in heavy metal concentration and exposure time had an adverse influence on the population growth of B. calyciflorus. However, while the response of B. calyciflorus was similar for both heavy metals, the magnitude of the impact of cadmium was more severe. Population growth varied depending on which heavy metal was tested, as well as its concentration and the duration of exposure (r = 0.11–0.28 day⁻¹). There was a significant reduction in lorica size of B. calyciflorus subjected to different exposure times and concentrations of both Cd and Cu. Egg ratios were inversely related to population density in controls and in treatments involving Cu, but not for Cd. While nearly 100% of eggs hatched in the control treatments, egg hatching in experimental treatments containing Cu, were reduced (range = 16–41%) depending on the exposure time and the concentration. Guest editors: S. S. S. Sarma, R. D. Gulati, R. L. Wallace, S. Nandini, H. J. Dumont & R. Rico-Martínez Advances in Rotifer Research     

Bemerkenswerte endogene Farbveränderung der Eischale von Chioninia delalandii (Duméril & Bibron, 1839) (Reptilia: Sauria: Scincidae)

The paper describes an unusual endogenous eggshell colouration observed in an egg of the Cap Verde skink Chioninia delalandii.A female specimen, kept in a terrarium, laid three eggs. Two of them were considered as fertilized (oval germ-disk, weakly pink). They were embedded 1 cm deep in a layer of moistened clay granules (substrate/water 2:1) and kept under different temperatures (egg 1 “cool”, 26-27 °C; egg 2 “warm”, 29-30 °C). There was a normal embryonic development in both eggs from their volume-enlargement and characteristic allometric growth (egg wide > egg length). The young hatch after 51 and 56 days.A change in eggshell colour, however, occurred in the “warm” kept egg during the last third of its incubation period. It started with a small spot (2-3 mm) in the outer area of the animal egg pole and spread into a dark-violet colouration over the whole eggshell within 15 days. After hatching of the young the shells of both eggs were examined. In the non-coloured egg there was no great difference in colour between the inner and outer egg layer, while in the coloured egg there was a distinct difference between the inner part, which was dark violet-gray, and the pale gray calcareous deck-layer. From the macroscopic view along the edge of the eggshell it was not identifiable, if the colour pigment was infiltrated into protein fibrils of the condensed surface layer.A possible explanation for the eggshell colouration could be an unusual embryonic pigmentation. This assumption is based on the first appearance of a restricted, point-like coloured area and its further regular extension. It might be that dark pigments (melanophores?) reached the eggshell (membrana testacea) and infiltrated the border-area to the condensed surface layer.     

Recovery of fat snook, Centropomus parallelus (Teleostei: Perciformes) after subchronic exposure to copper

We studied the recovery of juvenile fat snook (Centropomus parallelus) after subchronic exposure to different concentrations of copper. Healthy juveniles (1.98 g) were exposed to 25 or 50 μg Cu/L for 30 days (12 replicates with 5 fish in each one), and recovery was observed at 0, 4, 10, and 30 days after exposure (3 replicates with 5 fish in each one). Copper genotoxicity in exposed individuals was observed using a micronucleus assay, and recovery was not observed even 30 days post-exposure. Copper accumulation was observed in fish exposed to 25 or 50 μg/L of copper in the gills (14.4 and 34.4 μg/g, respectively) and muscle (5.7 and 5.5 μg/g, respectively), and a return to normal copper levels (6.0 μg/g for gills and 2.5 μg/g for muscle) was observed 4 and 30 days post-exposure in the gills and muscle tissues, respectively. Glutathione S-transferase (GST) was 80% inhibited in individuals exposed to copper and returned to normal levels for fish exposed to basal concentrations within 10 days. Although copper accumulation in tissues dispersed 30 days post-exposure, no recovery from genotoxicity was observed during this time. Thirty days was not enough to recover juvenile fat snook following subchronic exposure to copper.     

Cost of oviposition site selection in a water strider Aquarius paludum insularis: Egg mortality increases with oviposition depth

Females generally avoid selecting sites for oviposition which have a high predation risk to increase offspring survival. Previous studies have focused on costs to ovipositing females. However, although offspring may also incur costs by being oviposited at low predation risk sites, no studies have focused on costs to offspring. Such costs to offspring were examined by using Aquarius paludum insularis, females of which avoid eggs parasitism by ovipositing at deep sites. Deep sites are safe from egg parasitism but may be unsuitable for hatching due to environmental factors. We examined the costs to offspring at deep sites by comparing the hatching rate, the duration to hatching and the proportion of drowned larvae between eggs that were set at three levels of water depth (0 cm, 25 cm and 50 cm depth). While the hatching rate at 50 cm was lower than that at 0 cm, the rate at 25 cm did not differ from that at 0 cm. Duration to hatching and the proportion of drowned larvae did not differ between the three depths. It is suggested that the declining survival rate of A. paludum eggs was due to increased water pressure at greater depth. Such a cost may exist in other species and such an observation may aid in understanding oviposition site selection.     

Tributyltin inhibits the oligomycin-sensitive Mg-ATPase activity in Mytilus galloprovincialis digestive gland mitochondria

Tributyltin (TBT), widely employed in the past in antifouling paints, is one of the most toxic organic pollutants. Although recently banned, it still threatens coastal water ecosystems and accumulates in filter-feeding molluscs. TBT is known to act as a membrane-active toxicant; however data on mussels are scanty and exposure effects on mitochondrial ATPase activities remain hitherto unexplored. TBT effects on the mitochondrial Mg-ATPase activities in the digestive gland of Mytilus galloprovincialis were investigated both in vitro and in TBT-exposed mussels. Both an oligomycin-sensitive Mg-ATPase (OS Mg-ATPase) (70% of total Mg-ATPase activity) and an oligomycin-insensitive ATPase (OI Mg-ATPase) (30%) were found. The OS-Mg-ATPase was as much as 70% in vitro inhibited by 0.7 μM (203 μg/L) TBT, while higher concentrations promoted a partial inhibition release up to 5.0 μM TBT; higher than 10.0 μM TBT concentrations yielded nearly complete enzyme inhibition. Concentrations higher than 1 μM TBT enhanced the OI Mg-ATPase. Mussels exposed to 0.5 and 1.0 μg/L TBT in aquaria showed a 30% depressed OS Mg-ATPase activity, irrespective of TBT dose and exposure time (24 and 120 h). The OI Mg-ATPase activity was apparently refractory to TBT exposure and halved both in control and TBT-exposed mussels after 120 h exposure.     

Laboratory evaluation of the chemosterilant lufenuron against the fruit flies Ceratitis capitata, Bactrocera dorsalis, B. cucurbitae, and B. latifrons

Four species of tephritid fruit flies, Ceratitis capitata, Bactrocera dorsalis, B. cucurbitae, and B. latifrons were evaluated for toxic, developmental, and physiological responses to the chemosterilant lufenuron. No significant mortality of laboratory strains of the first three species was observed after their exposure up to 50 μg/mL of lufenuron in agar adult diet, whereas B. latifrons adults fed with 50 μg/mL of lufenuron in the diet caused significant mortality compared to the control. Fertility of C. capitata adults fed on 50 μg/mL lufenuron-fortified diet between 7 and 12 days of age was approximately 46% of the no lufenuron control. Fertility of B. dorsalis and B. latifrons adults fed on 50 μg/mL lufenuron-incorporated diet was about 45% and 62% of the control, respectively. Lufenuron did not significantly affect fertility of B. cucurbitae adults. Lufenuron did not affect fecundity of C. capitata and B. dorsalis. Fecundity of B. cucurbitae and B. latifrons was not evaluated due to difficulty to count the eggs laid deep in the agar diet. Larvae fed on a liquid larval diet with ≤ 0.1 μg/mL of lufenuron were also evaluated. Pupal recovery, adult emergence, adult fliers, mating, egg hatch, and egg production of C. capitata were significantly decreased, while for B. dorsalis, pupal recovery, larval duration and adult emergence were affected. No effect of lufenuron on B. cucurbitae larvae was observed. B. latifrons was not performed because shortage of eggs at the time of this research. Lufenuron is a potential agent for management and control of C. capitata and B. dorsalis.     

Cadmium uptake by rainbow trout, Salmo gairdneri eggs and alevins

The uptake of cadmium by eggs and alevins of rainbow trout from water concentrations of between 0.01 and 50.0 mg Cd 1⁻¹ was investigated. The cadmium content of eggs and alevins increased with time and with exposure concentration. Lower cadmium levels were detected in alevins than in eggs. Most of the cadmium (98%) in the eggs was found to be associated with the egg membrane or chorion. This explains the considerable reduction in cadmium concentration observed in alevins after hatching. Alevins hatching from eggs which had been exposed to cadmium survived longer in cadmium than alevins not exposed as eggs. This suggests that the pretreatment of eggs with cadmium serves some protective function. Behavioural and pathological signs of cadmium poisoning such as erratic swimming and blood clotting in alevins were observed.     

Metal accumulation and sublethal effects in the sea anemone, Aiptasia pallida, after waterborne exposure to metal mixtures

The marine environment is subjected to contamination by a complex mixture of metals from various anthropogenic sources. Measuring the biological responses of organisms to a complex mixture of metals allows for examination of metal-specific responses in an environmentally realistic exposure scenario. To address this issue, the sea anemone, Aiptasia pallida was exposed to a control and a metal mixture (copper, zinc, nickel, and cadmium) at three exposure levels (10, 50, and 100 μg/L) for 7 days. Anemones were then transferred to metal-free seawater for an additional 7 days after the metal exposure to assess metal depuration and recovery. Metal accumulation, activity of the enzymes catalase, glutathione reductase, and carbonic anhydrase, as well as, cell density of the symbiotic zooxanthellae were measured over 14 days. Metal accumulation in A. pallida occurred in a concentration dependent manner over the 7-day exposure period. Altered enzyme activity and tentacle retraction of the host, as well as decreased zooxanthellae cell density were observed responses over the 7 days, after exposure to a metal concentration as low as 10 μg/L. Metal depuration and physiological recovery were dependent on both the metal and the exposure concentration. Understanding how A. pallida and their symbionts are affected by metal exposures in the laboratory may allow better understanding about the responses of symbiotic cnidarians in metal polluted aquatic environments.     

Lavandula angustifolia essential oil as a novel and promising natural candidate for tick (Rhipicephalus (Boophilus) annulatus) control

Lavandula angustifolia is a well known herbal medicine with a variety of useful properties, including its acaricidal effect. This experiment was carried out to study the bioacaricidal activity of L. angustifolia essential oil (EO) against engorged Rhipicephalus (Boophilus) annulatus (Acari; Ixodidae) females. For this purpose six serial concentrations (0.5, 1.0, 2.0, 4.0, 6.0 and 8.0% w/v) of L. angustifolia EO were used. There was considerable mortality in concentrations more than 4.0% although there were no differences between 6.0 and 8.0% in all measured criteria. The mortality rate 24 h after inoculation was 73.26 and 100% in groups treated with 4.0 and 8.0% EO, respectively. Lavender EO also reduced tick egg weight in a concentration-dependent manner. The amount of eggs produced varied from 0.12 g (at 0.5% EO) to 0.00 g (at 8.0% EO) but did not differ statistically from the control. L. angustifolia EO caused 100% failure in egg laying at 6.0 and 8.0% whereas this value in the control group was zero. A positive correlation between L. angustifolia EO concentration and tick control, assessed by relative mortality rate and egg-laying weight, was observed by the EO LC/EC₅₀, which, when calculated using the Probit test, was 2.76-fold higher than the control. Lavender is a promising acaricidal against R. (B.) annulatusin vitro.     

The reproductive output of sea lice Caligus rogercresseyi under controlled conditions

Gravid females of Caligus rogercresseyi were collected from Atlantic salmon (Salmo salar) from a farm located at Chiloe Island (42°40′S73°15′W), Chile, to obtain information about the reproductive output of this parasite in vitro. The egg strings removed from the females were incubated under controlled conditions to obtain virgin adult females. One female which had mated only once produced eleven generations of eggs strings in a period of 74 days. The first egg strings of the females obtained in vitro were produced at 389 degree days (°D) after egg incubation, while the next generations of eggs strings were produced with a periodicity between 4 and 6 days dependent on the water temperature. The average length of the egg string was 3.1 mm and the mean number of eggs per string was 31. The values recorded in captivity for the egg string length and the number of eggs per string, were lower than the values recorded in gravid females from the field. One female survived for 79 days and males, maintained separately from the females, survived for 60 days.     

Influence of Temperature on Multiplication and Egg Hatching of Longidorus africanus

Longidorus africanus multiplication on tomato was highest at 29 °C. Few nematodes were recovered after 6 weeks at soil temperatures of 35 °C or below 23 °C. The time to egg hatching was shortest and the percentage of eggs hatching was highest at 29 °C. The minimum temperature and the heat sum above this temperature required for egg development were calculated to be 14.3 °C and 94.08 degree-days, respectively. The thermal times required for egg development by L. africanus and L. elongatus were nearly identical. For both species the product of the base temperature and the heat sum was near constant, and at a temperature of 22.3 °C the rates of egg development were equal.     

Leishmania (Viannia) peruviana (MHOM/PE/LCA08): Comparison of THP-1 cell and murine macrophage susceptibility to axenic amastigotes for the screening of leishmanicidal compounds

This study, undertaken to compare the susceptibility of THP-1 cells and murine peritoneal macrophages to Leishmania peruviana amastigotes, obtained THP-1 infection with 10 parasites/cell compared to 2 parasites/murine macrophage. The parasite burden was maximal at 72 h post-infection (h.p.i.) for THP-1 cells, while it was still increasing at 120 h.p.i. for murine macrophages. Since in both cases the infection with L. peruviana affected cell viability, we recommend evaluating any leishmanicidal activity at 72 h.p.i. Amphotericin B reduced Leishmania infection by 50% at concentrations of 0.1 μM in THP-1 and murine macrophages at 72 h.p.i.Our results demonstrate that amastigotes of L. peruviana can infect THP-1 cells and murine macrophages and indicate the suitability of this model to screen compounds for leishmanicidal activity.