Role of volatile semiochemicals in host location by the egg parasitoid Anagrus breviphragma

Recent investigations conducted on several tritrophic systems have demonstrated that egg parasitoids, when searching for host eggs, may exploit plant synomones that have been induced as a consequence of host oviposition. In this article we show that, in a system characterized by host eggs embedded in the plant tissue, naïve females of the egg parasitoid Anagrus breviphragma Soyka (Hymenoptera: Mymaridae) responded in a Y‐tube olfactometer to volatiles from leaves of Carex riparia Curtis (Cyperaceae) containing eggs of one of its hosts, Cicadella viridis (L.) (Hemiptera: Cicadellidae). The wasp did not respond to host eggs or to clean leaves from non‐infested plants compared with clean air, whereas it showed a strong preference for the olfactometer arm containing volatiles of leaves with embedded host eggs, compared with the arm containing volatiles of leaves from a non‐infested plant or host eggs extracted from the plant. When the eggs were removed from an infested leaf, the parasitoid preference was observed only if eggs were added aside, suggesting a synergistic effect of a local plant synomone and an egg kairomone. The parasitoid also responded to clean leaves from an egg‐infested plant when compared with leaves from a non‐infested plant, indicating a systemic effect of volatile induction.     

Host Adaptation and the Alteration of Viral Properties of the First Influenza A/H1N1pdm09 Virus Isolated in Japan

A/Narita/1/2009 (A/N) was the first H1N1 virus from the 2009 pandemic (H1pdm) to be isolated in Japan. To better understand and predict the possible development of this virus strain, the effect of passaging A/N was investigated in Madin-Darby canine kidney cells, chicken eggs and mice. A/N that had been continuously passaged in cells, eggs, or mice obtained the ability to grow efficiently in each host. Moreover, A/N grown in mice had both a high level of pathogenicity in mice and an increased growth rate in cells and eggs. Changes in growth and pathogenicity were accompanied by amino acid substitutions in viral hemagglutinin (HA) and PB2. In addition, the adapted viruses exhibited a reduced ability to react with ferret antisera against A/N. In conclusion, prolonged passaging allowed influenza A/N to adapt to different hosts, as indicated by a high increase in proliferative capacity that was accompanied by an antigenic alteration leading to amino acid substitutions.     

Dinitrocresol and phosphate stimulation of the oxygen consumption of a cell-free oxidative system obtained from sea urchin eggs

1. A cell-free system capable of using oxygen with oxalacetate as substrate has been prepared from both unfertilized and fertilized sea urchin eggs. The oxygen uptake by this system is about twice that of an equivalent quantity of intact unfertilized eggs and half that of an equivalent quantity of intact fertilized eggs. 2. The oxygen consumption of this cell-free oxidative system can be stimulated by addition of suitable concentrations of 4,6-dinitro-o-cresol or by inorganic phosphate. This confirms, with a cell-free system obtained from sea urchin eggs, the observations of Loomis and Lipmann regarding stimulation of oxygen consumption by a system obtained from rabbit kidney. 3. A preliminary but unsuccessful attempt has been made to determine the conditions under which cell-free, aerobic, phosphorylating systems may be obtained from either unfertilized or fertilized sea urchin eggs.     

Eggs survive through avian guts—A possible mechanism for transoceanic dispersal of flightless weevils

How flightless animals disperse to remote oceanic islands is a key unresolved question in biogeography. The flightless Pachyrhynchus weevils represent repetitive colonization history in West Pacific islands, which attracted our interests about how some weevils have successfully dispersed in the reverse direction against the sea current. Here, we propose endozoochory as a possible mechanism that the eggs of the weevils might be carried by embedded in the fruits as the food of frugivorous birds. In this study, Pachyrhynchus eggs were embedded in small pieces of persimmon fruits (Diospyros kaki) and fed to captive frugivorous birds. After digestion, 83%–100% eggs were retrieved from the feces of a bulbul (Hypsipetes leucocephalus) and two thrushes (Turdus chrysolaus). The retrieved eggs had hatching rates higher than 84%, which were not different from the control. In contrast, no egg was retrieved from the feces of the frugivorous pigeon (Treron sieboldii), which took a longer retention time in the guts. Our study identified that the eggs of Pachyrhynchus weevils are possible to be transported by internal digesting in some bird species.     

Cytological effects of urethan on cleavage induction in parthenogenetically activated sea urchin eggs

The effect of urethan on artificial induction of cleavage in eggs of the sea urchin, Hemicentrotus pulcherrimus, was studied. When the eggs were exposed for 20 minutes to seawater containing urethan (final concentration, 0.08 M) after butyric acid-activation and then treated with hypertonic seawater, the cleavage rate was enchanced by about 1.5 times as compared with the nontreated eggs. In the eggs exposed to urethan–seawater for over 70 minutes many clear spots appeared throughout the cytoplasm. Simultaneously, the pigment granules, which had been embedded within the cortex, migrated to the inner cytoplasm and encircled a monastral centrosphere and clear spots. The clear spots were composed of microtubules much like cytasters, and in the central region of them centrioles were not yet found. The eggs in which the pigment granules disappeared from the cortex may be more susceptible to cleavage induction by succeeding hypertonic treatment.     

Angiostrongylus vasorum infection in a coyote (Canis latrans) from Newfoundland and Labrador, Canada

Tissue samples and feces were collected from a dead, adult female coyote (Canis latrans) found at the side of the road in late March 2003 in the Avalon Peninsula region of Newfoundland, Canada. The coyote apparently died of vehicular-related trauma. Samples of lung, brain, heart, liver, and kidney were fixed in formalin and submitted for histologic examination. The entire remaining lung and heart also were submitted for examination. The coyote was diagnosed with moderate, multifocal, granulomatous interstitial pneumonia with eosinophilic vasculitis and many intralesional nematode eggs, larvae, and occasional intravascular adult worms. Adult nematodes recovered from the pulmonary arteries were identified as Angiostrongylus vasorum. Small foci of granulomatous inflammation, often containing nematode eggs and larvae, were scattered in the brain and kidney. To our knowledge, this is the first report of A. vasorum infection in a coyote from the only endemic area of infection in North America.     

The spatial distribution of maternal mRNA is determined by a cortical cytoskeletal domain in Chaetopterus eggs

Messenger RNA molecules are localized in the cortical region of eggs and unevenly segregated to the embryonic cells during early development of the annelid Chaetopterus. The egg cortex is enriched in two organelles, ectoplasmic spherules and associated structures, which are similar in appearance to nuage. The physical basis of cortical mRNA localization was examined in stratified eggs and in eggs extracted with the nonionic detergent Nonidet P-40 (NP-40). The cortical organelles were displaced to the most centrifugal zone of stratified eggs. In situ hybridization with poly(U) or cloned DNA probes showed that a large proportion of the poly(A)+RNA, histone mRNA, and actin mRNA molecules was also displaced to the centrifugal zone. Extraction with NP-40 revealed a detergent-insoluble cytoskeletal domain (CD) in the egg cortex which contained the remnants of ectoplasmic spherules and nuage embedded in a fibrous network. Although most of the total protein and RNA was extracted by NP-40, a large proportion of the poly(A)+RNA, histone mRNA, and actin mRNA molecules was retained in the CD. In situ hybridization of stratified eggs extracted with NP-40 indicated that the CD, with its associated organelles and mRNA molecules, is displaced to the centrifugal zone as a unit. The results suggest that the tenacious association of mRNA molecules with the cortical CD may be responsible for maternal mRNA localization during early development.     

Learedius learedi infection in black turtles (Chelonia mydas agassizii), Baja California Sur, Mexico

Black turtle (Chelonia mydas agassizii) carcasses, recovered as a result of incidental capture in Magdalena Bay, Mexico, revealed invasion by spirorchiid trematode eggs in liver, kidney, intestines, muscle, heart, pancreas, and duodenum. Seventy-five adult Learedius learedi Price, 1934, were recovered from the heart of 1 turtle. Most of the organs showed a mild or absent inflammatory response in histological sections, with the exception of a pancreatic-duodenal section that revealed severe lymphocyte and phagocyte infiltration associated with an infestation of more than 200 eggs. A linear formation of 35 eggs from the pancreas toward the intestinal lumen is described as resembling migration. This is among the first reports of a parasitic infection of L. learedi Price 1934, in C. m. agassizii in Mexico.     

转基因红鲤体细胞的核移植

以F4代转hGH基因红鲤体细胞（肾脏和尾鳍）及培养18代的F4代转hGH基因红鲤尾鳍细胞为核供体,泥鳅或黄河鲤成熟卵为受体,进行了核移植,以探讨外源F4代转基因鱼体外源基因的分布与存在形式,稳定性和克隆转基因鱼的可能性。F4代红鲁肾脏细胞核与泥鳅卵配合的核移植胚胎有12．4％发育到囊胚,0．33％发育到神经胚;F4代尾鳍细胞核移入泥鳅卵后的重组胚发育到囊胚,神经胚、肌节期和肌肉效应期的胚胎分别为24．5％、0．3％、0．2％和0．1％;对照卵无发育。F4代红鲤尾鳍培养细胞与黄河鲤卵子配合的重组胚胎有50．53％发育到囊胚,5．69％发育到原肠胚,0．53％发育到神经胚,0．4％发育到肌节期。说明由于同种细胞核与卵细胞的相容性高于异种核卵的相容性,早期发育率高;而由于培养细胞的异倍化,后期的发育率降低。用PCR技术对供体鱼不同个体及同一体不同组织外源基因检测,结果100％个体为阳性鱼,而且不同组织的阳性率也是100％,说明外源基因均匀分布在不同组织中。无论F4代转基因鱼的肾脏细胞、尾鳍细胞还是培养的尾鳍细胞作核移植供体,核移植胚胎中hGH基因的检出率为100％。说明F4代转基因红鲤个体不同细胞都存在hGH基因,而且经长期培养不会丢失。表明F4代转基因红鲤中的外源hGH基因已基本稳定,体细胞核移植可以作为获得同质化转基因鱼的有效手段,但核移植效率还很低。另外还讨论了核质的相容性、细胞周期的协调、染色体的变异等因素对核移植的影响。     

Acceptance and suitability of four plant substrates for rearing Orius sauteri (Hemiptera: Anthocoridae)

Orius sauteri (Poppius) is an important hemipterous predator that has been mass-reared for biological control of numerous pests in protected crop-production systems. To find a good oviposition substrate for mass-rearing this predator under insectary conditions [25°C, 65 ± 5% relative humidity, and a photoperiod of 16:8 (L:D) h], we compared kidney bean, soybean, broad bean sprouts and fresh leaves of kidney bean. We found that O. sauteri made more punctures and laid more eggs in kidney bean sprouts than in the other substrates examined. However, there were no significant differences among substrates in the proportion of punctures receiving eggs. Female O. sauteri laid the most eggs (as many as 68 eggs) in kidney bean sprouts and also had the shortest pre-oviposition period on this plant material. In addition, there were no significant differences in total oviposition durations or female longevity among the four plant substrates. The hatch rates of nymphs in the sprouts and leaves of kidney bean (>90%) were higher than those in soybean and broad bean sprouts. Thus, we found that the kidney bean sprout was the most suitable substrate for mass-rearing of O. sauteri.     

Pluripotency of Homozygous-Diploid Mouse Embryos in Chimeras

Pluripotency of mouse uniparental cells (complete homozygous-diploid gynogenetic) produced by embryo manipulation was examined in aggregation chimeras with normally fertilized embryos. A male pronucleus was removed from fertilized eggs by micromanipulation and eggs were diploidized with cytochalasin B. Uniparental cells that developed to 4-cell or more advanced stages were aggregated with normally fertilized 8-cell embryos and transferred to the pseudopregnant female uteri to develop to term. Among the pups, 1 female and 3 males were identified as overt chimeras by their coat color and pigmentation of the retina. Using electophoretic analysis of the isozymes, the contribution of uniparental cells in these chimeras was confirmed by findings in the major organs such as liver, brain, small intestine, kidney, spleen, heart and testis. The female chimera produced offspring derived from oocytes of uniparental origin. Our experiments verified the pluripotency of microsurgically produced mouse uniparental cells.     

Cytostatic factor (CSF) activity in cytosols extracted from Xenopus laevis eggs

Cytostatic factor (CSF), found in the cytoplasm of unfertilized eggs of amphibians, causes metaphase arrest when microinjected into cleaving blastomeres. Although CSF from Rana pipiens eggs has been extracted and characterized, little is known about CSF extracted from eggs of other species. We investigated the conditions required to preserve CSF activity in cytosols extracted from Xenopus laevis eggs and found that it was necessary to expose the eggs to CO2 prior to extraction and that the extraction buffer must contain sodium beta-glycerophosphate. CSF activity disappeared after 24 h of storage at 2 degrees C. Cytological examination showed that the arrested blastomeres injected with cytosols had been arrested at metaphase and contained a spindle lacking polar asters, in which highly condensed chromosomes were embedded.     

Modèle de cage permettant d'obtenir la ponte d'un diptèreBombyliidae,villa quinquefasciata Wied. AP. Meig.

Summary This paper describes a breeding cage for the adults ofVilla quinquefasciata. The cage (3×4×2 meters high; wooden frame supporting a plastic screen) lies out of doors. Twenty centimeters above the ground, there is a wooden floor pierced by a number of holes. Somes of these receive boxes filled with sand; in the others, are embedded wooden panels provided with a circular opening of about 20 centimeters in diameter. A wire screen is fixed under each panel and bears one or several stones. Finally, a removable funnel is fixed under each panel and is fitted into a vial. The females fill up their ?perivaginal pouch? in the sand boxes, and then eject the eggs towards the base of the stones. The eggs, as they fall, are collected inside the vials. Nevertheless, as the wooden frame casts a shadow, the females eject also their eggs all around the cage. These eggs are collected using a small vacuumcleaner. The adults mate easily in captivity. They refuse to feed, and their longevity is low (no more than 6 days). This cage allows one, however, to easily obtain a large number of eggs from a rather small number of adults (for example, in 1965, 42,000 eggs from 503 adults).      

Quantification of spermatozoa in the sperm-storage tubules of turkey hens and the relation to sperm numbers in the perivitelline layer of eggs

This study was conducted to determine the number of spermatozoa residing in the oviduct sperm-storage tubules (SST) and the relationship between these numbers and the number of spermatozoa embedded in the perivitelline layer of oviductal eggs after a single insemination of 200 x 10(6) spermatozoa. The SST of hens inseminated within one week before the expected onset of egg production were filled faster (4 h vs. 2 days) and possessed more spermatozoa (4.1 vs. 2.0 x 10(6)) than the SST of hens inseminated after the onset of egg production. Furthermore, for hens in egg production, significantly fewer spermatozoa were recovered from the SST if the hen was inseminated within 2 h before or after oviposition than if inseminated more than 2 h before or after the oviposition. There was a strong positive correlation between the number of spermatozoa in the SST and the number of spermatozoa embedded in the perivitelline layer of the oviductal eggs (r = 0.85, p less than 0.01). These data show that the population of spermatozoa actually accepted by the SST is quite small relative to the number of spermatozoa inseminated and that maximum sperm-storage is achieved when the hen is inseminated just prior to the onset of egg production. It is suggested that the sperm-storage capacity of the oviduct and the quality of the semen sample can be estimated on the basis of numbers of spermatozoa embedded in the egg perivitelline layer.     

Studies on enzymes of nitrogen metabolism, and nitrogen end products in the developing chick (Gallus domesticus) embryo.

1. A total of 450 fertilized eggs were used to study the concentrations of uric acid, urea and ammonia in allantoic and amniotic fluids, and some enzymes of nitrogen metabolism in the liver and kidney during the development of the chick embryo from the 5th to 21st day of incubation. 2. Concentrations of the compounds studied were higher in allantoic fluid. The molar concentration of allantoic uric acid increased steadily with time. The pattern of urea and ammonia in both allantoic and amniotic fluids were the same. 3. Arginase (E.C.3.5.3.1) activity in both embryonic kidney and definitive kidney was higher than that in the liver. The specific activity of arginase (mumole urea formed/hr per g wet wt kidney) dropped during development. 4. Little arginine synthetase activity (argininosuccinate synthetase, E.C.6.3.4.5; and argininosuccinate lyase, E.C.4.3.2.1) was found in kidney, but none in the liver. 5. The complete urea cycle function was absent in both the liver and the kidney of the chick embryo.     

Concentrations of erythromycin and azithromycin in mature Chinook salmon Oncorhynchus tshawytscha after intraperitoneal injection, and in their progeny

A single dose (40 mg kg(-1)) of erythromycin or azithromycin dihydrate was injected intraperitoneally into maturing female fall Chinook salmon 12 to 32 d before spawning to observe the distribution, retention and clearance of the drugs in plasma, kidney, coelomic fluid and egg vitellin, and their persistence in alevins derived from these fish. Salmon administered prophylactic dosages of erythromycin as subadults were also included to investigate potential interactive effects of oral and injected treatments on reproductive performance and antibiotic clearance. Erythromycin was rapidly cleared from plasma and coelomic fluid, but was detected in the kidney (3.52 to 12.40 microg g(-1)) and egg vitellin (5.32 to 8.87 microg ml(-1)) of all fish at spawning. High, stable concentrations of azithromycin were detected in plasma (14.66 to 20.33 microg ml(-1)), kidney (43.16 to 59.96 microg g(-1)), coelomic fluid (2.52 to 5.50 microg ml(-1)) and egg vitellin (12.65 to 23.51 microg ml(-1)). Oral administration of erythromycin to subadult salmon did not significantly affect tissue concentrations of either erythromycin or azithromycin administered by prespawning injection. Reductions in the percentage of eggs that yielded live embryos at the eyed stage of development occurred among eggs derived from females that had received orally administered erythromycin as subadults. Erythromycin was not detected in unfed fry derived from adults injected with the drug prespawning, but azithromycin was present for more than 2 mo after the onset of exogenous feeding.     

Pathogenicity of avian nephritis virus for embryonating hen's eggs

The pathogenicity of avian nephritis virus (ANV) for embryonating hen's eggs was studied by various routes of inoculation. When inoculated with ANV by the yolk sac route, 6-day-old embryos showed the highest susceptibility and all of them died 3 to 14 days postinoculation (PI). They manifested hemorrhage and edema of the whole body (3 to 6 days PI) and stunting (7 to 14 days PI). The 50% egg-infective dose of the virus by yolk sac inoculation coincided well with the virus titer expressed in plaque-forming units determined on the monolayer of chicken kidney cell cultures. The virus could be passed serially through the chorioallantoic membrane (CAM) of embryonating hen's eggs. In these eggs the CAM presented edematous thickening at the inoculation site, and the embryo stunting. when inoculated by the CAM route, high virus doses killed all embryos, but low virus doses allowed some of the infected embryos to hatch normally. When inoculated by the allantoic cavity route, the virus did not multiply in the allantoic cavity of embryonating eggs, but some of these eggs became infected. Fluorescent antigens were present only in the kidneys and CAM of embryos infected with the virus. The virus was recovered at a low rate from cloacal swabs of chicks from normally hatched eggs inoculated with the virus by the CAM route. These chicks were variable in growth, but had antibodies against the virus and developed nephritis at 36 days of age.     

Melamine in eggs,plasma and tissues of hens fed contaminated diets

A study was conducted to evaluate the excretion pattern of melamine from feed into eggs, plasma, kidney, liver and muscle of laying hens. In particular, 90 laying hens were randomly allocated to three dietary treatments and fed diets contaminated with melamine at a level of 2.5, 25 and 250 mg of melamine/kg of diet for T1, T2 and T3 groups, respectively. The diets were offered in six replicate boxes (five hens each) for 13 days. Eggs were collected from each group for melamine quantification on days 0, 1, 3, 6, 9 and 13. At the end of the experimental period, one hen per box was randomly selected and slaughtered to collect plasma, liver, kidney and muscle samples. During the experiment, feeding diets with increasing levels of melamine had no effect (P > 0.05) on weight gain, feed intake, egg production, egg weight and mortality of laying hens. The melamine in eggs increased from day 1 after melamine ingestion and reached a plateau between days 6 and 13 of melamine ingestion. At steady-state condition, the melamine egg concentrations increased (P < 0.01) with treatments, being 0.026, 0.352 and 4.631 mg/kg for T1, T2 and T3, respectively. Similarly, the carryover of melamine from feed to egg increased (P < 0.05) with the levels of melamine in the diets, varying from 0.50 to 0.70 and 0.84 for T1, T2 and T3, respectively. The melamine was detected in plasma of all tested groups, increasing (P < 0.01) with levels of melamine in the diets (0.030, 0.266 and 4.102 mg/l in T1, T2 and T3, respectively). Melamine was not detected in kidney, liver and muscle of hens fed T1. Except for kidney sampled in the T3, no melamine concentration higher than 2.5 mg/kg, representing the maximum allowable limit set by the US Food and Drug Administration and European Union for food and feeds, was measured. The melamine resulted higher in plasma and kidneys than in the liver and muscle both in T2 and T3. The results confirmed the presence of an excretion pattern of melamine from feed to eggs and tissues in laying hens.     

Physiological responses of sea urchin eggs to stimulation by calcium ionophore A23187 analysed with protease inhibitors

Protease inhibitors were used to study certain physiological responses (secretion of the cortical granule protease, altered resceptively to sperm penetration, initiation of cell division and embryogenesis) of sea urchin eggs to stimulation by calcium ionophore A23187. Protease activity in the secretory product released from the eggs 5 min after insemination or parthenogenetic activation with ionophore was completely inhibited by soybean trypsin inhibitor (SBTI), antipain (Ap), and leupeptin (Lp). A barrier was established to prevent subsequently added sperm from penetrating (fertilizing) ionophore-activated eggs, co-incident with the elevation of the fertilization membrane. These processes were retarded by inhibitors of the cortical granule protease in ionophore-activated eggs, just as they are when eggs are initially stimulated by sperm at fertilization. A23187-activated eggs did not divide unless they had been secondarily fertilized by sperm, even if the ionophore was subsequently removed by extensive washing. However, ionophore-activated eggs that were penetrated by a single spermatozoan in SBTI developed into normal larvae under similar conditions. These results suggest that A23187 may be an incomplete parthenogenetic agent because it cannot stimulate eggs to assemble centrioles required to organize the mitotic apparatus. The centrioles are normally provided by the sperm during fertilization. A23187 may also be toxic to the eggs. Furthermore, since cortical granules are secretory organelles, the data suggest a possible functional relationship between calcium ions and protease activation in stimulus-secretion coupling in sea urchin eggs at fertilization.