Ex situ transportation of coral larvae for research,conservation, and aquaculture

Due to the lack of appropriate methods to transport high amounts of larvae ex situ over large distances, the availability of coral larvae was so far mainly limited to their place of origin. For a research project at Rotterdam Zoo, The Netherlands, we transported several thousand larvae of three broadcast spawners (Acropora tenuis, A. digitifera, Diploria strigosa) from the Indo Pacific and the Caribbean to Europe. Beside logistics and packing techniques, post-transport survival rates were mainly influenced by larvae density and transport duration. Our results indicate optimum survival rates of >90% at densities of 4 larvae ml⁻¹ when not exceeding a transportation time of 4 days. The ex situ transport of coral larvae over large distances might offer new possibilities for research, conservation, and aquaculture.     

Diel timing of planulation and larval settlement in the coral Isopora cuneata (Scleractinia: Acroporidae)

The prevailing environmental conditions when reproductive products are released in the water column can have a profound influence on the biology and ecology of marine organisms. In reef-building corals, brooding species that release azooxanthellate larvae are expected to release their larvae before sunrise, similar to species releasing zooxanthellate larvae as established in previous studies. This study investigated the diel timing of release of asymbiotic larvae by Isopora cuneata in northwestern Philippines during 2 years through ex situ observations on gravid colonies collected from the reef. Planulation mainly occurred after sunset until midnight, with grand mean hour of release at 19:35 h (95% confidence limit: 18:44–21:02 h) in April 2010 and 19:41 h (18:10–20:59 h) in March 2011. This pattern of release timing is distinct from what has been previously reported for brooding scleractinian coral species with zooxanthellate larvae (i.e., release close to sunrise). The release coincides with the low tidal stand, which may provide opportunity for the prompt settlement of newly released, buoyant larvae to shallow reef areas where adult colonies are generally distributed. The larvae were able to settle with 20 ± 25% success within a day of release, though a distinct settlement peak at 3 days post-release (70 ± 25%). This study provides new information on the early life strategies of the reef-building coral Isopora cuneata. The unique pattern of larval release time and the settlement behavior may influence the population dynamics and success of the species through space and time.     

Mutual interference in the mass-reared fruit fly parasitoid,Diachasmimorpha longicaudata (Hymenoptera: Braconidae)

Diachasmimorpha longicaudata is a solitary endoparasitoid produced in Mexico for the biological control of Anastrepha fruit flies (Diptera: Tephritidae). We studied the effect of mutual interference among conspecific foraging females to better understand the parasitoid-host dynamics established in the mass-rearing system of this species. We used a constant host availability of 60 third instar larvae of Anastrepha ludens (per oviposition unit type Petri dish) that were individually exposed for 3 h to 1, 2, 4, 8, 10, 14, and 20 D. longicaudata females, seven days old without oviposition experience. The following parameters were evaluated: total number of attacked hosts (i.e., larvae with at least one oviposition scar), number of attacked hosts per female, adult emergence percentage, and female proportion. We also performed a second assay at mass-rearing level, contrasting the current proportion used in mass-rearing (~?two host larvae per female) with the alternative treatments (5 and 7.5 host larvae per female). Results showed that the density of females foraging on a patch exhibits an inverse relationship to the number of hosts attacked by one female. The highest values for adult emergence (73.4%), patch exploitation (94.6%), and female proportion (0.86) were obtained in the treatments with 7.5, 6, and 3 host larvae per female, respectively. Under mass-rearing conditions, we found that a lower density of females per cage (~?five larvae per female) reduced superparasitism levels without affecting adult emergence and the proportion of females. This suggests that the mass-rearing efficiency of D. longicaudata could be improved by reducing the density of foraging females.

     

Effects of growing resistant potatoes with gene H1 from Solarium tuberosum ssp. andigena on populations of Heterodera rostochiensis British pathotype A

Pure pathotype A populations of Heterodera rostochiensis produce a few females on ex andigena hybrids with the H₁ gene for resistance. As the proportion of larvae able to become female on ex andigena hybrids was not increased by reproducing the nematodes on such hybrids for 3 years, these females seem not to be genetically different from the rest of the population. The proportion increased rapidly when the initial population contained a few pathotype (species) E nematodes but again no increase in the proportion of pathotype (species) A larvae able to become female on ex andigena was detected and pathotype E replaced pathotype A.     

Comparison of the Host Searching and Oviposition Behaviors of the Tephritid (Diptera) Parasitoids Aganaspis pelleranoi and Odontosema anastrephae (Hymenoptera: Figitidae, Eucoilinae)

We compared the host-searching and oviposition behaviors of two Neotropical figitid parasitoids (Hymenoptera) that exploit the same resource: ripe fruit infested by fruit fly larvae (Tephritidae) that have fallen to the ground. Sexually mature Aganaspis pelleranoi (Brèthes) and Odontosema anastrephae Borgmeier females were exposed individually, under no choice conditions, to four types of fruit: 1) Clean, intact guavas, Psidium guajava L. (no fruit fly larvae, no perforations); 2) clean, with artificial perforations; 3) artificially infested (with larvae), no perforations; 4) infested with artificial perforations. A behavioral transition matrix and sequence diagram of the following behaviors was constructed: walking on fruit, detection of larvae via the antennae, tarsi or aculeus, fruit perforation and penetration, and oviposition. Overall, we found that infested fruit (intact and with artificial perforations) elicited the most activity in the females of both species and that A. pelleranoi females exhibited a significantly more diverse behavioral repertoire (i.e., more transitions) and were significantly more active than O. anastrephae females. Females of both species penetrated the fruit in search of larvae by biting through the epi- and mesocarp, but O. anastrephae remained inside for significantly longer periods (up to eight hours). A. pelleranoi females used both their antennae and tarsi to detect larvae but the use of these structures varied depending on context: in infested fruit tarsi were used preferentially (usually while standing still) while in uninfested fruit, antennae were mainly used (usually while walking). In the case of O. anastrephae females the reverse pattern was usually observed with antennae most commonly used to detect larvae in infested fruit. We discuss our findings in light of their evolutionary, ecological and practical implications.     

Seasonal dynamics of agonistic behavior and hormones in an ex situ all‐male colony of large flying foxes

Large flying foxes (Pteropus vampyrus) are a socially complex species. In situ colonies typically comprise thousands of individuals in small harems of one male to many females. In ex situ environments, all‐male colonies are becoming more common due to a surplus of males in the population. There is limited information describing the hormonal and behavioral patterns of all‐male colonies during the breeding season. We assessed seasonal changes in hormones and behavior in an all‐male colony of 12 large flying foxes at Disney's Animal Kingdom^®. We validated hormone assays using morning urine and fecal samples to assess seasonal changes in excreted immunoreactive testosterone and glucocorticoid metabolites. We collected behavior data using an all‐occurrence method, recording agonistic behaviors related to territorial defense (hooking, biting, wing flexing, vocalizing, and wrestling), and sexual behavior (mounting and frontal grabbing). Results indicated that (i) we could reliably measure testosterone and glucocorticoid metabolites concentrations from fecal and urine samples collected from individual bats; (ii) there were distinct relationships between changes in levels of agonism and hormone concentrations throughout the year; and (iii) three agonistic behaviors (chasing, wrestling, and open‐mouth threat) peaked prior to the increase in testosterone and glucocorticoid hormones measured during the breeding season. These three behaviors could potentially be used as early indicators to signal the onset of the breeding season and allow time to implement ex situ management changes to reduce the incidence of agonism between individuals.     

Reproductive and subsocial behaviour in the ovoviviparous leaf beetle Gonioctena sibirica (Coleoptera: Chrysomelidae)

Abstract.

• 1 In the spring, females of the leaf beetle Gonioctena sibirica deposited larvae on the ventral surface of growing young leaves situated on the apical position of shoots of the willow Salix bakko.
• 2 The parent females remained with the larvae usually on the underside of the basal part of leaves, facing toward the base of shoots. When other arthropods approached, the females temporarily moved towards these intruders, showing aggressive behaviour such as swinging the body or stamping the legs. Many females remained with their larvae until the larvae grew into the final (fourth) instar. No female produced an additional brood in the field.
• 3 Broods from which parent females were experimentally removed suffered higher mortality than those in which females were left intact. Arthropods such as spiders and ants were observed preying on the larvae. In contrast, the survivorship of broods from which females were removed and intruders were excluded with a sticky substance applied to the base of twigs was not different from that of control broods. These results demonstrate that the main mortality factor of offspring is pedestrian arthropod predators and females physically repel the predators.
• 4 Potentially alternative reproductive strategies, such as producing a large number of offspring by iteroparity and/or larger brood(s) with less or no care, seem to be inhibited in G.sibirica by larval dependence on growing young leaves which are temporally limited and by ovoviviparity which may have limited brood size.

     

Reproductive performance of clonal and sexual bark beetles (Coleoptera: Scolytidae) in the field

In Ips acuminatus (Gyll.) parthenogenetic females occur together with sexual females and with sexual males upon which they depend for sperm. In a reciprocal‐transplant experiment, I studied fecundity differences among parthenogenetic and sexual females from two populations that differ dramatically in the proportion of clonal females. In a second experiment, I studied competition between larvae from different mothers and between females from the two source populations. Fecundity measured by the number of eggs per egg tunnel was influenced by the ambient environment at the sites of the experiment as well as the origin of the female, and was generally higher for clonal than for sexual females at both sites. In experimental groups where larvae competed with larvae from their own population (pure treatments), the number of surviving pupae was significantly lower than in groups where females from the two source populations were mixed. The high fecundity of clonal females makes coexistence of the two types of females difficult to explain. It makes the reproductive advantage associated with clonality in I. acuminatus even higher than the two‐fold difference due to asexuality per sé. The significant differences in the number of pupae in mixed vs. pure groups suggest ecological divergence between sexual and clonal females. This would make the mortality of larvae not only density dependent, but also frequency dependent, which could explain the coexistence of sexual and clonal females.     

Clutch size in a larval-pupal endoparasitoid

Clutch size decisions by Aphaereta minuta (Nees) (Hymenoptera: Braconidae), a polyphagous, gregarious, larval-pupal endoparasitoid, were studied under laboratory conditions. This parasitoid attacks larvae of Diptera inhabiting ephemeral microhabitats such as decaying plant and animal material. Females oviposit in young larval stages, but the eventual size of the host pupa determines host food availability for competing offspring. The size of the pupa can differ greatly between host species. We questioned how A. minuta females deal with this delay between the moment of oviposition and eventual host food availability, and whether they make clutch size decisions that benefit their fitness. It was shown that females indeed vary their clutch size considerably and in an adaptive way: (1) females lay larger clutches in larvae of host species that produce larger pupae, even when the larvae are the same size at the moment of oviposition, and (2) females lay larger clutches in larger larvae than in smaller larvae of the same host species. The latter seems functional as larvae parasitized at an older stage indeed developed into larger pupae compared to larvae parasitized at a younger stage. Furthermore, mortality of parasitized young host larvae was greater than that of both unparasitized larvae and parasitized older larvae. Under field conditions the risk of mortality of young host larvae is expected to be even higher due to the limited period of microhabitat (host food) availability, strong scramble type competition between the host larvae, and the longer period of being exposed to predation.     

Ovipositional discrimination by Microplitis rufiventris females between healthy and granulosis virus-infected Spodoptera littoralis larvae

Ovipositional choice tests by Microplitis rufiventris females (Hym., Braconidae) between granulosis virus‐infected (GVI) and non‐infected (NI) Spodoptera littoralis larvae (Lep., Noctuidae), were assessed using discriminatory methods for re‐isolating the NI and virus‐infected hosts after removing the female parasitoid. When M. rufiventris females were given a choice between NI and GVI S. littoralis hosts, the adult females exhibited marked preference (P < 0.01) for the NI (i.e. higher quality) hosts. In this case, M. rufiventris females and S. littoralis GV (SlGV) did not significantly compete for the same type of host larvae and are, generally, compatible. However, when the choice was given between two low qualities of S. littoralis hosts, i.e. virus‐free previously parasitized hosts and viral‐infected hosts a significant preference (P < 0.01) of the parasitoid females for the GVI larvae was observed. In this case, the parasitoid would be at a disadvantage when competing with GV for the same host. However, the parasitoid could be used as an additional tool for the dissemination of biocontrol viruses within different pest populations, i.e. hosts other than S. littoralis. Importantly, the results showed different strategies of parasitoid female in egg‐laying management. When M. rufiventris female was given a choice between healthy and SlGVI hosts, the female deposited more eggs than when she was given a choice between two low qualities of host larvae. The results of the study may have implications in pest management strategies using M. rufiventris and SlGV against S. littoralis larvae.     

The Andean bear alopecia syndrome may be caused by social housing

The Andean bear alopecia syndrome is a progressive and chronic condition documented in ex situ populations. Recent advances focus on treating symptoms, not preventing future cases. We therefore explored the epidemiology of this syndrome through an analysis of husbandry and veterinary conditions of 63 Andean bears (26M:37F) housed in North and South American zoos and other ex situ circumstances. We had the most complete information for the North American population and found that 29% of females (n = 24) were affected. No males (n = 26) were affected. An analysis of generalized linear models indicated that three models were competitive in describing the occurrence of the condition (i.e., ΔAIC_c ≤ 2): the model including only the individual's sex (χ² = 13.41, df = 1, p < .001), the model including both individual sex and social housing status (χ² = 1.36, df = 2, p < .001), and the model including both individual sex and the expression of stereotypical behaviors (χ² = 13.82, df = 2, p = .001). Stereotypical behaviors were common among both males (50%, n = 26) and females (51.9%, n = 27) whether or not they were affected, but the syndrome was seen only in females who had been socially housed. Therefore, we suggest that the Andean bear alopecia syndrome is a symptomatic response to the long‐term social housing of bears that would otherwise not live socially. To prevent new cases, we recommend that female Andean bears be housed with adult conspecifics only when females choose to cohabitate.     

The egg size determining gene,Esd, is a unique morphological marker on the W chromosome ofBombyx mori

Triploid females of the silkworm,Bombyx mori, were produced by crossing a tetraploid female with a male carrying asch (sex-linked chocolate) gene on its Z chromosomes. Since thesch gene expresses the color of reddish brown (chocolate) in newly hatched larvae, the females with ZWW chromosomes become chocolate, while those with ZZW have normal color (black). The effects of the W chromosome on oogenesis were studied by comparing these two types of 3n females. Despite the fact that the theoretical value of the ratio of chocolate larvae to black larvae should be 1:5, the actual ratio was found to be 1:80. The chocolate larvae revealed very low hatchability. Thirteen adult ZWW female moths were obtained, all laying normal-shaped eggs and even larger than those of 4n females which also have two W chromosomes. Eighty percent of the eggs of ZZW 3n females were irregular-shaped, and the size of the normal-shaped eggs was as large as that of 2n females which possess one W chromosome. It is hypothesized that egg size is determined by a gene on the W chromosome. The higher frequency of normal-shaped eggs in the ZWW female group than the ZZW female group suggests the presence of another kind of gene on the W chromosome, which leads to normal oogenesis.     

Host discrimination and superparasitism in wild and mass-reared Diachasmimorpha longicaudata (Hym.: Braconidae) females

We compared the oviposition behavior and host discrimination ability of wild and mass-reared Diachasmimorpha longicaudata (Hymenoptera: Braconidae) females parasitizing Anastrepha ludens (Diptera: Tephritidae) larvae. Both kinds of parasitoid females were presented simultaneously with parasitized and non-parasitized larvae in choice tests, and their superparasitism performance was evaluated under a mass-rearing situation. At the time of the test, D. longicaudata had 156 generations under mass-rearing conditions. Our goal was to determine the effect of the mass-rearing process on the foraging decisions of this species. One of the primary findings was the apparent ubiquity of superparasitism by D. longicaudata females. Both types of females showed similar patterns in each of the phases of oviposition behavior evaluated. The only notable differences were among the percentages of transition between behaviors, mainly related to the intensity with which each activity was performed. Under a mass-rearing situation, both strains of females had a similar tendency to increase superparasitism (i.e., number of oviposition scars per puparium and the proportion of superparasitized larvae) over time. The mass-rearing process appears to have induced the selection of more aggressive, fertile and precocious females. Despite these observations, we concluded that the process of adaptation to mass-rearing conditions has not substantially influenced the foraging and ovipositional behaviors in this species.     

Changes in the chromosomal insertion pattern of the copia element during the process of making chromosomes homozygous in Drosophila melanogaster

In situ hybridization on polytene chromosomes of Drosophila melanogaster was used to compare the insertion patterns of copia and mdgl transposable elements on chromosome 2 in male gametes sampled by two different methods: (i) by crossing the males tested with females from a highly inbred line with known copia and mdgl insertion profiles; (ii) by crossing the same males with females from a marked strain, and analysing the resulting homozygous chromosomes. Crossing of the males with the inbred line led to homogeneous insertion profiles for both the copia and mdgl elements in larvae, thus giving an accurate estimation of the patterns in the two gamete classes of each male. Crossing with the marked strain led, however, to heterogeneity in insertion patterns of the copia transposable element, while no significant polymorphism was observed for mdgl. The use of balancer chromosomes is thus not an adequate way of inferring transposable element insertion patterns of Drosophila males, at least for the copia element. This technique could, however, be powerful for investigating the control of movements of this element.     

Host selection by the autoparasitoid Encarsia pergandiella on primary (Bemisia tabaci) and secondary (Eretmocerus mundus) hosts

In autoparasitoids, females are generally primary endoparasitoids of Hemiptera, while males are hyperparasitoids developing in or on conspecific females or other primary parasitoids. Female‐host acceptance can be influenced by extrinsic and/or intrinsic factors. In this paper, we are concerned with intrinsic factors such as nutritional status, mating status, etc. We observed the behavior of Encarsia pergandiella Howard (Hymenoptera: Aphelinidae) females when parasitizing primary (3rd instar larvae of Bemisia tabaci Gennadius [Homoptera: Aleyrodidae]) and secondary hosts (3rd instar larvae and pupae of Eretmocerus mundus Mercet [Hymenoptera: Aphelinidae]) for a period of 1 h. Females had different reproductive (virgin or mated younger) and physiological (fed elder or mated elder) status. Virgin females killed a large number of secondary hosts while investing a long time per host. However, they did not feed upon them. Mated females killed a lower number of secondary hosts and host feeding was observed in both consuming primary and secondary hosts. It was common to observe host examining females of all physiological statues tested repeatedly stinging the same hosts when parasitizing, killing or rejecting them. Fed elder females parasitized more B. tabaci larvae than E. mundus larvae or pupae, while investing less time on the primary host than on the secondary host. They also parasitized more B. tabaci larvae than mated elder females, while investing less time per host. The access of females to honey allowed them to lay more eggs.     

Taste sensitivity and divergence in host plant acceptance between adult females and larvae of Papilio hospiton

On the island of Sardinia the lepidopteran Papilio hospiton uses Ferula communis as exclusive host plant. However, on the small island of Tavolara, adult females lay eggs on Seseli tortuosum, a plant confined to the island. When raised in captivity on Seseli only few larvae grew beyond the first–second instar. Host specificity of lepidopterans is determined by female oviposition preferences, but also by larval food acceptance, and adult and larval taste sensitivity may be related to host selection in both cases. Aim of this work was: (i) to study the taste sensitivity of larvae and ovipositing females to saps of Ferula and Seseli; (ii) to cross‐compare the spike activity of gustatory receptor neurons (GRNs) to both taste stimuli; (iii) to evaluate the discriminating capability between the two saps and determine which neural code/s is/are used. The results show that: (i) the spike responses of the tarsal GRNs of adult females to both plant saps are not different and therefore they cannot discriminate the two plants; (ii) larval L‐lat GRN shows a higher activity in response to Seseli than Ferula, while the opposite occurs for the phagostimulant neurons, and larvae may discriminate between the two saps by means of multiple neural codes; (iii) the number of eggs laid on the two plants is the same, but the larval growth performance is better on Ferula than Seseli. Taste sensitivity differences may explain the absence of a positive relationship between oviposition preferences by adult females and plant acceptance and growth performance by larvae.     

potamolejeunea Holtii (Spruce) nov.comb.

Abstract

Bryophytes usually have anti-feeding properties to defend against microbial and herbivore attack; however, the consumption of Haplocladium microphyllum (Hedw.) Broth. capsules by Agrotis sp. larvae is rather common in Shanghai in the spring. To test whether H. microphyllum is the only moss eaten, and why the gametophytes of H. microphyllum are not eaten, a series of quantitative experiments were carried out to understand the feeding habits of Agrotis larvae on the given moss materials at three growth stages of larval life. The results show that the larvae can feed on the capsules of six moss species to different degrees: Funaria hygrometrica Hedw., H. microphyllum, Physcomitrium sphaericum (C.F.Ludw. ex Schkuhr) Brid., Trematodon longicollis Michx., Ditrichum pallidum (Hedw.) Hampe, and Pogonatum inflexum (Lindb.) Sande Lac. The capsules of the first four species were grazed heavily by the larvae, compared with limited consumption of the latter two (D. pallidum and P. inflexum), which even induced a high mortality rate among the larvae. With the growth of the larval instar, the daily demand for moss capsules by the larvae increases gradually. The lipid content of the capsules may play an important role when the larva selects its feeding target.     

The effect of resistant solanaceous plants on the sex ratio of Heterodera rostochiensis and the use of the sex ratio to assess the frequency and genetic constitution of pathotypes

The sex ratios of two Heterodera rostochiensis populations were compared on fifteen solanaceous plants. Larvae did not mature in the roots of Atropa belladona, Datura stramonium or Nicotiana tabacum but adults were produced on the other plants, on some only males and on others, males and females. With two other populations, but not with a third, the percentage of larvae that became female on a potato hybrid with a dominant gene for resistance derived from Solanum tuberosum ssp. andigena increased during successive generations on the hybrid in accordance with the hypothesis that such larvae are double recessives (aa). Fewer larvae became adult in plants with two dominant genes for resistance than in plants with one, and such plants probably exerted selection of both males and females. A population from St Brelades (Jersey) produced fewer females and more males on the susceptible variety Arran Banner than did either the Thorney (Cambs.) or Aberystwyth populations. A much greater percentage of larvae from the St Brelades population became female on an ex andigena hybrid derived from CPC 1690 than on one derived from CPC 1685.     

Hybridization between Helicoverpa armigera and Helicoverpa assulta (Lepidoptera: Noctuidae): development and morphological characterization of F1 hybrids

Reciprocal hybridizations between Helicoverpa armigera (Hübner) and Helicoverpa assulta (Guenée) were studied. The cross between females of H. armigera and males of H. assulta yielded only fertile males and sterile individuals lacking an aedeagus, valva or ostium bursae. A total of 492 larvae of the F1 generation were obtained and 374 of these completed larval development and pupated. Only 203 pupae were morphologically normal males, the remaining 171 pupae were malformed. Larvae and pupae that gave rise to morphologically abnormal adults exhibited longer development times. Sterility was not only associated with malformed external sex organs, but also a range of abnormalities of the internal reproductive system: (i) loss of internal reproductive organs, (ii) with one to three copies of an undeveloped bursa copulatrix; or (iii) with one or two undeveloped testes. Normal male hybrid adults showed higher flight activity in comparison with males of both species. In contrast, the cross between females of H. assulta and males of H. armigera yielded morphologically normal offspring (80 males and 83 females). The interaction of the Z-chromosome from H. assulta with autosomes from H. armigera might result in morphological abnormalities found in hybrids and backcrosses, and maternal-zygotic incompatibilities might contribute to sex bias attributed to hybrid inviability.     

Genetic variation and conservation assessment of Chinese populations of Magnolia cathcartii (Magnoliaceae), a rare evergreen tree from the South-Central China hotspot in the Eastern Himalayas

Nine natural populations of the rare evergreen tree Magnolia cathcartii (Magnoliaceae) were sampled across its natural range, and amplified fragment length polymorphism (AFLP) markers were used to assess genetic variation within and among populations. Three ex situ populations were also surveyed to determine whether conservation plantings include the entire genetic diversity of the species. Genetic diversity within the natural populations was very low (0.122 for Nei’s gene diversity), and the southeast populations had the highest diversity. The ex situ populations had a lower diversity than the mean diversity for all populations, and none of the ex situ populations reached the levels of diversity found in their source populations. Genetic differentiation was high among natural populations (G _st = 0.247), and an isolation-by-distance pattern was detected. Habitat fragmentation, restricted gene flow, and geitonogamy are proposed to be the primary reasons for the low genetic diversity and high genetic differentiation. More protection is needed, especially for the southeast populations, which possess the highest numbers of unique alleles according to AFLP fragment analyses. The ex situ program was a good first step towards preserving this species, but the current ex situ populations preserve only a limited portion of its genetic diversity. Future ex situ efforts should focus on enhancing the plantings with individuals from southeastern Yunnan.