Detection of only JH III in several life-stages of Nauphoeta cinerea and Thermobia domestica

We have conducted a reinvestigation into both the identification and quantification of juvenile hormone (JH) from several developmental stages of the cockroach, Nauphoeta cinerea, and the firebrat, Thermobia domestica, using a gas chromatography-mass spectrometric (GC/MS) method. We detected only JH III in these animals in contrast to prior studies in which JH I, II, and/or III had been reported using a different scheme relying on HPLC purification and subsequent GC/MS analysis under chemical ionization (CI) conditions. Very high levels (approximately 800 ng/g) of JH III were found in N. cinerea embryos at stages after dorsal closure whereas first stadium nymphs and female penultimate stadium nymphs contained only low levels (approximately 1 ng/g and approximately 7 ng/ml respectively); in adult females at the stage of rapid oocyte growth approximately 150 ng JH III per ml of hemolymph was measured. T. domestica nymphs and egg laying adults contained only low levels (approximately 1 ng/g) of JH III. The results emphasize the caution which must be used in interpreting results of procedures for analysis of JH at parts-per-billion levels, and also enforce prior observations that the higher JH homologs are not present except in the Lepidoptera.     

Factors affecting survival of mouse blastocysts vitrified by a mixture of ethylene glycol and dimethyl sulfoxide

The present study was conducted to determine suitable conditions for mouse blastocysts vitrified by a solution containing 25 % v/v (4.5M) ethylene glycol and 25% v/v (3.4M) dimethyl sulfoxide (VSi). In Experiment 1, blastocysts were exposed to 50% diluted VSi (50% VSi) for 10 minutes then to VSi for 0.5 minutes at room temperature (22 approximately 24 degrees C) or at 4 degrees C, followed by vitrification. The survival rates of these embryos exposed at each temperature were not significantly different. In Experiment 2, embryos were exposed directly to VSi for various time periods at room temperature and diluted in mPBS with 0.5 M sucrose without vitrification. The viability of embryos decreased after more than a 3 minute exposure. When the embryos were exposed to VSi for 0.5, 1, 1.5 and 2 minutes followed by vitrification, the survival rates were 78, 80, 76 and 50%, respectively. In Experiment 3, embryos were vitrified after exposure to 50% VSi for various time periods and then to VSi for 0.5 minutes at room temperature. One minute exposure to 50% VSi resulted in the highest survival rate. In Experiment 4 and 5, the cooling rate (from approximately 70 to approximately 2500 degrees C/minute), sucrose concentration (0, 0.5 and 1 M) of dilution solution, and dilution time (1 or 5 minutes) did not affect the viability of the vitrified embryos. Following exposure to 50% VSi for 1 minute and to VSi for 0.5 minutes at room temperature, embryos were cooled 1) at approximately 2500 degrees C/minute and diluted in 0.5 M sucrose in mPBS after warming or 2) at approximately 200 degrees C/minute and diluted in mPBS. In vivo development rates after the transfer to recipients were 38 and 42%, respectively. These values were similar to that of fresh control embryos.     

Mdogg1基因参与家蝇体内氧化还原平衡的维持

[目的]本研究旨在克隆鉴定家蝇Musca domestica中8-羟基鸟嘌吟糖苷酶1(8-hydroxyguanine glycosylase 1,OGG1)编码基因Mdogg1,明确其是否参与家蝇氧化应激调控.[方法]根据家蝇转录组数据,利用RT-PCR克隆Mdogg1基因cDNA序列,并进行生物信息学分析;利用qR...     

Severely reduced recombination in females of the South American marsupial Monodelphis domestica.

The gray short-tailed opossum, Monodelphis domestica, is a model species for studying the underlying mechanisms that govern recombination. We have identified a new marker, PI (protease inhibitor), in M. domestica and demonstrate its linkage to AK1 (adenylate kinase 1). The rate of recombination in females of this species is approximately one fifth the rate in males, as might be predicted from the difference in chiasma frequency between the two sexes.     

Nymphal Development and Reproduction of Podisus nigrispinus (Heteroptera: Pentatomidae) Fed With Combinations of Tenebrio molitor (Coleoptera: Tenebrionidae) Pupae and Musca domestica (Diptera: Muscidae) Larvae

The development and reproduction of Podisus nigrispinus (Dallas) (Heteroptera: Pentatomidae) fed on Tenebrio molitor L. (Coleoptera: Tenebrionidae) and Musca domestica L. (Diptera: Muscidae) were studied in four treatments. P. nigrispinus was permitted to feed on: (1) T. molitor pupae (T 1 ); (2) Musca domestica larvae (T 2 ); (3) both prey supplied simultaneously (T 3 ); (4) both prey supplied on alternate days (T 4 ). Duration of the nymphal period of P. nigrispinus was similar in all diets studied with nymphal viability of approximately 75%. Heavier females were obtained in T 1 and T 4 , but no correlation between this factor and the reproductive rate of the predator was found. Therefore, the use of body weight as a parameter to evaluate rearing quality should be approached with caution. However, females of this predator showed higher egg and nymph production when they received both prey. For this reason P. nigrispinus should be reared with T. molitor and M. domestica simultaneously or on alternate days.     

Thermogenic capabilities of the opossum Monodelphis domestica when warm and cold acclimated: similarities between American and Australian marsupials

1. Monodelphis domestica is a small marsupial mammal from South America. Its thermogenic abilities in the cold were determined when the opossums were both warm (WA) and cold (CA) acclimated. Maximum heat production of M. domestica was obtained at low temperatures in helium-oxygen. 2. Basal metabolic rate (BMR) in the WA animals was 3.2 W/kg and mean body temperature was 32.6 degrees C at 30 degrees C. These values were lower than those generally reported for marsupials. Nevertheless, these M. domestica showed considerable metabolic expansibility in response to cold. Sustained (summit) metabolism was 8-9 times BMR, while peak metabolism was 11-13 times BMR. These maximum values were equal to, or above, those expected in small placentals. 3. Cold acclimation altered the thermal responses of M. domestica, particularly in warm TaS. However, summit metabolism was not significantly increased; nor did M. domestica show a significant thermogenic response to noradrenaline, which in many small placentals elicits non-shivering thermogenesis. The thermoregulatory responses of this American marsupial were, in most aspects, similar to those of Australian marsupials. This suggests that the considerable thermoregulatory abilities of marsupials are of some antiquity.     

Comparative toxicity of seven insecticides to immature stages of Musca domestica (Diptera: Muscidae) and two of its important biological control agents, Muscidifurax raptor and Spalangia cameroni (Hymenoptera: Pteromalidae)

The toxicity of seven insecticides was evaluated against unparasitized Musca domestica L. pupae and pupae parasitized by Muscidifurax raptor Girault & Sanders or Spalangia cameroni Perkins, two important biological control agents. Only pyrethrins + piperonyl butoxide (Pyrenone) was less toxic to M. raptor compared with house flies. Conversely, all of the insecticides except crotoxyphos were less toxic to S. cameroni compared with house flies. A plateau in the tetrachlorvinphos bioassay line for S. cameroni suggested that this colony had approximately 45% resistant individuals. The selectivity observed between immature stages of house flies and M. raptor or S. cameroni is different from that reported against adult stages of these same species, suggesting that selectivity of an insecticide varies considerably between different life stages.     

Photorepair of ultraviolet radiation-induced pyrimidine dimers in corneal DNA

The induction and photorepair of pyrimidine dimers in DNA have been measured in the ultraviolet-irradiated, corneal epithelium of the marsupial, Monodelphis domestica, using damage-specific nucleases from Micrococcus luteus in conjunction with agarose gel electrophoresis. We observed that FS-40 sunlamps (280-400 nm) induced 7.2 +/- 1.0 X 10(-5) pyrimidine dimers per kilobase (kb) of DNA per J/m2. Following 100 J/m2, 50% and greater than 90% of the dimers were photorepaired during a 10- and 30-min exposure to photoreactivating light (320-400 nm), respectively. In addition, approximately 70% and approximately 60% of the dimers induced by 300 and 500 J/m2, respectively, were repaired by a 60-min exposure to photoreactivating light. The capacity of the corneal epithelium of M. domestica to photorepair pyrimidine dimers identifies this animal as a potentially useful model with which to determine whether pyrimidine dimers are involved in pathological changes of the irradiated eye.     

Foraging behavior of the queenless ant Dinoponera quadriceps Santschi (Hymenoptera: Formicidae)

The search for and ingestion of food are essential to all animals, which spend most of their lives looking for nutritional sources, more than other activities such as mating, intra-specific disputes or escaping from predators. The present study aims to describe and quantify several aspects of foraging behavior, diet and food transport in the queenless ant Dinoponera quadriceps Santschi in a secondary Atlantic forest, Northeastern Brazil. Three colonies were randomly selected at a distance of at least 50 m from one another. On leaving the colony, worker ants were followed until their return, with no nutritional provision or interference with their activities. Activities were recorded using focal time sampling with instantaneous recording every minute for 10 consecutive minutes. Each colony was observed 1 day/week, for at least 6 h/day resulting in 53.8h of direct observation of the workers. Foraging activities, success in transporting food, type of food, cleaning and interaction among the workers were recorded. Foraging was always individual, with no occurrence of recruitment. Diet was composed mainly of arthropods, mostly insects. The collection of small fruits (Eugenia sp.) was also observed. Foraging time was greater when workers transported food to the colony, the return time being shorter than the foraging period, suggesting the use of chemical and visual cues for orientation during their foraging and food-collecting activities.     

Nature of the erythroid colony stimulating extract obtained from spleen of irradiated rats (author's transl)]

Non-adherent bone marrow cells of a bled rabbit were cultured in plasma clot media containing auto-serum, alpha-medium, erythropoietin (Ep) and spleen extract from irradiated rats. The preparations were cloted on a cover glass, fixed and stained by Giemsa or hemoglobin staining method after 3 or 5 days in culture, and the number of erythroid colony was counted as reported elsewhere. In the present study, first, it was elucidated that the optimal numbers of innoculating cells were among 0.6 approximately 1.2 x 10(4) cells per well for the erythroid colony formation. Second, this colony formation was slightly stimulated by the experimental media which contained heat treated extract at 40 degree or 50 degree C for 30 minutes. Contrary this, the extract treated at 70 degree C for 30 minutes lost completely its stimulating activity of the colony formation, suggesting that the effective substances might be protein in the extract. Third, an inhibitory factory might be present in the dialysate of the crude spleen extract, because the number of erythroid colonies decreased in a dose response manner by the dialysate. The residue of inner dialysate, however, certainly contained the colony stimulating factors (s). The crude extract was separated into five fractions (F1 approximately F5) by ammonium sulfate. F1, which was precipitated with 40% ammonium sulfate, had the highest activity for the colony formation. Fetuin also showed appreciable effect on the erythroid colony formation.     

Methodological studies on growth of mouse granulocyte/macrophage colonies in methylcellulose-containing media

We studied the influence of various physicochemical parameters on colony development and total cell numbers in 7-day methylcellulose cultures of mouse bone marrow cells. Colony growth was markedly retarded by an increase of PO2 from approximately 6.7 kPa towards that in ambient air and by a decrease of incubator temperature from 37 degrees C to 33 degrees C. Medium osmolality above approximately 340 mosm/kg inhibited formation of granulocytes (in cultures containing growth factors from pokeweed mitogen-stimulated spleen cells), but not macrophages (L cell-produced growth factors). At most, colony growth was affected slightly by moderate changes in pH (7.17-7.47) or concentration of methylcellulose (0.77-1.02%), or by the presence of 2-mercaptoethanol (50 mumol/1), Hepes buffer (25 mmol/1), or erythropoietin (0.1-1 units/ml). The culture trays could be stored for one day at 4 degrees C in the incubation boxes before colonies or cells were counted.     

Musca domestica is not a vector of Thelazia callipaeda in experimental or natural conditions

Thelazia callipaeda Railliet and Henry (Spirurida: Thelaziidae), commonly called oriental eyeworm for its widespread presence in the Far East, has been recently found to affect dogs, cats and foxes in northern and southern Italy. Although the biology of T. callipaeda in the definitive hosts has been recently investigated, many doubts still remain about its biology in insect vectors. It has been suggested that more than one species of Diptera, namely Musca domestica Linnaeus (Diptera: Muscidae) and Amiota okadai Maca (Diptera: Drosophilidae), may be involved in the transmission of T. callipaeda in China. The aim of the work described here was to verify the role of M. domestica as a vector of T. callipaeda both in experimental and natural conditions. A total of 310 m. domestica (Group 1) were put in a cage and allowed to feed for 14 days around the eyes of a dog naturally infected by T. callipaeda. Ten flies were collected daily for 14 days. A total of 149 houseflies (Group 2) were fed with T. callipaeda first stage larvae (L1) and dissected at 1, 2 and 7 days post-infection. From June to August 2003, flies were netted (Group 3) in two different sites every 10 days both from the environment and directly from the periocular region of dogs affected by thelaziosis. Musca flies were examined for eyeworms by dissection and visual inspection of house flies (Groups 1 and 2) and using a molecular approach (Groups 1-3) via a specific amplification of the ribosomal internal transcribed spacer 1 (ITS1) sequence of T. callipaeda. On the whole, 180 pools of M. domestica flies were processed molecularly and all the experimentally infected flies (Groups 1 and 2) were found to be negative both at the visual dissection and at the molecular assay. Similarly, the 234 M. domestica collected from Group 3 were negative for T. callipaeda. The results clearly suggest that M. domestica is unlikely to act as a vector of T. callipaeda in southern Europe, in contrast with a single previous report.     

Detection, isolation and some properties of membrane proteinases from yeast mitochondria

The complete amino-acid sequences of two isocytochromes c (larval- and adult-type cytochromes c) purified from the housefly Musca domestica L. were determined. Their sequences differed at six positions from each other. More than 90% of the total cytochrome c was larval-type during larval stages. The amount of adult-type cytochrome c increased rapidly from 1 day before adult emergence, making the total cytochrome c content increase to approximately 2.5-times as much cytochrome c content as in larvae. Although the major cytochrome c species in flight muscles was adult-type, imaginal disks contained mainly larval-type cytochrome c.     

Genetic diversity of laboratory gray short-tailed opossums (Monodelphis domestica): effect of newly introduced wild-caught animals.

The colony of gray, short-tailed opossums (Monodelphis domestica) at the Southwest Foundation for Biomedical Research, the primary supplier of this species for research purposes, was founded with nine animals trapped in 1978 in the state of Pernambuco, Brazil. Since 1984, 14 newly acquired founders from the state of Paraiba, Brazil have contributed to the gene pool of the colony. The animals from Paraiba and their descendants are significantly larger than the founders from Pernambuco and their descendants. The two groups also differ significantly in several measurements of morphologic traits. The changes in proportional contribution of each founder to the colony, and changes in inbreeding coefficients during the colony's history, are evaluated. Using previously established markers and three newly identified markers (ACP2, APRT, and DIA1), we show that the Paraiba-derived animals differ significantly from the original founders in allele frequencies and heterozygosity. The genetic diversity of the colony has been substantially increased by acquisition of the new founders from Paraiba. The colony is highly polymorphic, with 22.2% of loci surveyed by protein electrophoresis being variable. We conclude that the genetic differences between populations and among projects within the colony should be considered in future colony management procedures and in selection of experimental subjects.     

家蝇卵黄蛋白基因的克隆与结构序列分析

家蝇卵黄蛋白基因编码的卵黄蛋白是家蝇胚胎发育的重要营养来源 .根据 3种家蝇卵黄蛋白cDNA保守序列设计引物 ,用PCR技术从家蝇基因组DNA中扩增到大小为 76 8bp的mdYP1基因的部分DNA片段 .经地高辛标记成特异性探针 ,从构建的家蝇基因组文库中筛选出一个阳性克隆 ,并从该克隆中分离到大小为 3991bp的mdYP1基因组基因 .序列分析显示 ,该基因组序列含有约1 6kb的 5′ 上游区和 1 0kb的 3′ 下游区 ,编码区由一个 6 1bp的内含子和大小分别为 2 2 2bp和10 2 8bp的 2个外显子组成 .5′ 上游区含有典型的CAAT TATA盒 .     

The Azolla, Anabaena azollae Relationship: II. Localization of Nitrogenase Activity as Assayed by Acetylene Reduction

Anaerobic (microaerophilic) acetylene reduction by Azolla caroliniana Willd. was dependent on light and saturated at approximately 450 foot candles. Maximum rates of acetylene reduction were 60 nmoles/mg chlorophyll minute. However, rates of 25 to 30 nmoles/mg chlorophyll minute were more common.     

Linkage mapping and physical localization of the major histocompatibility complex region of the marsupial Monodelphis domestica

We used genetic linkage mapping and fluorescence in situ hybridization (FISH) to conduct the first analysis of genic organization and chromosome localization of the major histocompatibility complex (MHC) of a marsupial, the gray, short-tailed opossum Monodelphis domestica. Family based linkage analyses of two M. domestica MHC Class I genes (UA1, UG) and three MHC Class II genes (DAB, DMA, and DMB) revealed that these genes were tightly linked and positioned in the central region of linkage group 3 (LG3). This cluster of MHC genes was physically mapped to the centromeric region of chromosome 2q by FISH using a BAC clone containing the UA1 gene. An interesting finding from the linkage analyses is that sex-specific recombination rates were virtually identical within the MHC region. This stands in stark contrast to the genome-wide situation, wherein males exhibit approximately twice as much recombination as females, and could have evolutionary implications for maintaining equality between males and females in the ability to generate haplotype diversity in this region. These analyses also showed that three non-MHC genes that flank the MHC region on human chromosome 6, myelin oligodendrocyte glycoprotein (MOG), bone morphogenetic protein 6 (BMP6), and prolactin (PRL), are split among two separate linkage groups (chromosomes) in M. domestica. Comparative analysis with eight other vertebrate species suggests strong conservation of the BMP6-PRL synteny among birds and mammals, although the BMP6-PRL-MHC-ME1 synteny is not conserved.     

Laboratory and field evaluation of formulated Bacillus thuringiensis var. israelensis as a feed additive and using topical applications for control of Musca domestica (Diptera: Muscidae) larvae in caged-poultry manure

Infestations of house flies, Musca domestica L., are a continual problem around poultry establishments. Acute toxicity of two commercial Bacillus thuringiensis variety israelensis (Bti) formulations (water-dispersible granules and bran formulation) was evaluated against larvae in the laboratory and against natural populations of M. domestica larvae in the field applied in feed to chickens and as topical applications in the poultry houses. Bioassay data showed that susceptibility of M. domestica larvae increased to a given concentration of Bti as the duration of exposure increased. In the laboratory studies, the LC(50) values of Bti for the larvae ranged between 65 and 77.4 μg/ml. In the field, a concentration of 10 g Bti/kg of feed resulted in 90% reduction of larvae at 4 wk after treatment. A higher concentration (2 g/liter) of Bti in spray applications was not significantly more effective than the lower concentration of 1 g/liter. Adding Bti to chicken feed is potentially an efficient measure for the management and control of house flies in caged-poultry facilities.     

Neurotensin regulation of macrophage colony-stimulating factor-stimulated in vitro myelopoiesis

Neurotensin, at less than or equal to 10(-9) M, in the presence of an optimal concentration of macrophage CSF (CSF-1), stimulated a dose-dependent enhancement of colony formation by murine marrow-derived mononuclear phagocyte progenitor cells. The additional colonies arose from the cell cycle and Ia Ag-positive subpopulation previously identified as two-signal-dependent progenitors. Two-signal colony formation diminished when the peptide was added at concentrations greater than 10(-9) M. Neurotensin binds specifically to two distinct receptors, a high affinity receptor (KD approximately 10(-9) M) and a lower affinity (KD approximately 10(-7) M) receptor identified as the tuftsin receptor. Rat liver ferritin and an inhibitory tuftsin analog. (ALA1)-tuftsin, which inhibit two-signal colony formation stimulated by tuftsin and tuftsin-like peptides in combination with CSF-1, did not inhibit colony formation stimulated by CSF-1 and 10(-9) M neurotensin. Both inhibitors, however, reversed the loss of two-signal colony growth in the presence of higher neurotensin concentrations. Neurotensin fragment 1-6, unlike ferritin and (ALA1)-tuftsin, inhibited two-signal colony formation stimulated by 10(-9) M neurotensin. However, like ferritin and (ALA1)-tuftsin, fragment 1-6 permitted full expression of two-signal colony formation in the presence of CSF-1 and 10(-7) M neurotensin. The data indicated that occupancy of both receptors at neurotensin concentrations greater than 10(-9) M might be responsible for the diminished progenitor response. The data further support a potential role for neurotensin as an inflammatory mediator. In addition to direct effects on mature phagocytic leukocytes, neurotensin, at least in vitro can influence the production of new mononuclear phagocytes.     

An evaluation of (Z)-9-tricosene and food odours for attracting house flies, Musca domestica, to baited targets in deep-pit poultry units

Field trials investigating the effect of food baits on catches of Musca domestica at toxic targets impregnated with the female sex pheromone, (Z)-9-tricosene, were conducted in a caged-layer deep-pit poultry unit in southern England. Targets treated with an Alfacron-sugar mixture and baited with 2.5 g of 40% (Z)-9-tricosene beads caught significantly greater numbers of both male and female M. domestica than control targets. Egg and milk-baited targets were less attractive than controls, while brewers yeast slightly increased the numbers of M. domestica attracted. However, the inclusion of brewers yeast in (Z)-9-tricosene-impregnated targets produced a significant reduction in the number of male M. domestica attracted. Increased female attraction was elicited by baiting the targets with 2-phenylethanol, at the quantities of 1 mg and 10 mg. However, 2-phenylethanol had no effect on female attraction when presented in conjunction with (Z)-9-tricosene. The implications of these results in relation to the control of M. domestica populations in poultry units are discussed.