Efficacy of oral sulfadimethoxine against two gregarine parasites,Protomagalhaensia granulosae and Gregarina cubensis (Apicomplexa: Eugregarinida), infecting the Death's Head cockroach,Blaberus discoidalis

Gregarines are common parasites of insects in culture, but no effective chemotherapeutic or prophylactic control protocol has been demonstrated. Sulfadimethoxine was administered in 5- and 7-day treatments to Death's Head cockroaches (Blaberus discoidalis) infected with Gregarina cubensis and Protomagalhaensia granulosae to test the efficacy of this sulfonamide against gregarine infection. Sulfadimethoxine significantly reduced the mean intensity of both G. cubensis and P. granulosae. Sulfadimethoxine treatment reduced gregarine intensity by 80% to 85% but had no significant effect on gametocyst production, suggesting that sulfonamide toxicity is directed primarily at sporozoites, trophozoites, and perhaps young gamonts. The possible use of sulfadimethoxine to produce gregarine-free insect cultures and the potential utility of gregarines as target organisms for screening pharmacologically active compounds for use against other intestinal apicomplexans are discussed.     

Effects of pH on Excystation of Gregarina cuneata and Gregarina polymorpha (Eugregarinida: Gregarinidae)1

ABSTRACT. The mechanisms that signal gregarine excystation are unknown. Previous authors have demonstrated that gregarine sporozoites excyst from their surrounding oocyst in response to stimuli contained in host digestive fluids, but the role of host intestinal pH in this signaling system has not been investigated. In this study, an in vitro assay is used to quantify the effects of 3 pH levels (6.1, 7.0, 8.0) on the excystation of two gregarine species, Gregarina cuneata and Gregarina polymorpha. Both gregarine species excyst at all three pH levels, but there are significant within and among species differences in excystation rate and cumulative excystation over time. Gregarina cuneata excysts more rapidly at pH 6.0 and G. polymorpha excysts more rapidly at pH 8.0. Cumulative excystation is maximized at pH 6.0 for G. cuneata and at pH 7.0 for G. polymorpha. Hydrogen ion mediated excystation may lead to the formation of foci for subsequent establishment or migration and may play a role in parasite site specificity.     

Genetic control of Eucalyptus urophylla and E. grandis resistance to canker caused by Chrysoporthe cubensis

Chrysophorte cubensis induced canker occurs in nearly all tropical and subtropical regions where eucalypts are planted, causing losses in both wood quality and volume productivity, especially so in the warmer and more humid regions of Brazil. The wide inter and intra-specific genetic variability of resistance to canker among Eucalyptus species facilitates the selection of resistant plants. In this study, we evaluated resistance to this pathogen in five Eucalyptus grandis (G) and 15 E. urophylla (U) trees, as well as in 495 individuals from 27 progenies derived from crosses between the trees. In the field, six-months-old test seedlings were inoculated with C. cubensis. Lesion length in the xylem and bark was measured eight months later. The results demonstrated that xylem lesions could preferentially be used for the selection of resistant clones. Eight trees (7 U and 1 G) were susceptible, and the remainder (8 U and 4 G) resistant. Individual narrow and broad sense heritability estimates were 17 and 81%, respectively, thereby suggesting that canker resistance is quantitative and highly dependent on dominance and epistasis.     

Amoebogregarina Nigra n. gen., n. comb. (Apicomplexa: Gregarinidae) from adult Melanoplus differentialis (Orthoptera: Acrididae) in southeastern Nebraska

Amoebogregarina nigra n. gen., nov. comb. (Eugregarinida: Gregarinidae) is described from trophozoites, gamonts, associations, gametocysts, and oocysts collected from adult Melanoplus differentialis (Orthoptera: Acrididae) in Nemaha County, Nebraska. Host alimentary canals were examined for eugregarine parasites. Gregarines encountered were fixed as permanent specimens or subjected to a series of morphometric measurements. Morphometric analysis indicated the presence of Gregarina nigra, a poorly described taxon reported from a variety of Nearetic grasshopper species, Examination of G. nigra revealed a metamorphic epimerite assimilated by the protomerite on maturity. This epimerite-protomerite complex is unique within Gregarinidae, prompting creation of the genus Amoebogregarina. Amoebogregarina nigra is the type species in new combination. Gregarina indianensis is recognized as a junior synonym of A. nigra.     

高温诱导黄瓜抗霜霉病机理

研究了高温对黄瓜霜霉病菌致病力的影响以及高温控制霜霉病发生的效果.结果表明,40 ℃高温处理2 h和45 ℃处理1 h对黄瓜霜霉病的诱导抗病性作用最明显,其在接种后4 d时的防效分别为58.40%和45.81%,到接种后6 d时,防效分别下降为39.35%和37.65%.经高温诱导后,过氧化物酶（POD）、苯丙氨酸解氨酶(PAL)、几丁质酶（Cht）、β-1,3-葡聚糖酶（Glu）活性均显著高于对照,与未诱导植株相比,高温诱导后叶片组织的细胞壁表面有大量木质素沉积,表明高温处理后黄瓜表现出对霜霉病的抗病性.     

The life cycle of Gregarina ronderosi n. sp. (Apicomplexa: Gregarinidae) in the Argentine grasshopper Dichroplus elongatus (Orthoptera: Acrididae)

Gregarina ronderosi n. sp. is described based on life cycle observations conducted on nymphs and adults of its natural host, the grasshopper Dichroplus elongatus. Following ingestion of oocysts by the host, parasite development occurs between the epithelium and the food mass in the midgut and gastric caeca. Gametocysts are liberated in the faeces. Natural prevalence in the type locality, Girondo, northwestern Buenos Aires Province, was 39.7% (n=131). The earliest trophozoites seen were small (< or = 10 microm), somewhat ovoid, unsegmented bodies. Fully developed trophozoites (the body is divided into epimerite, protomerite, and deutomerite) were slender, with conical or globular epimerites in attached or unattached forms, respectively. Trophozoites varied greatly in size [total length: 10.4-275.1 microm; mean (+/-S.E.): 126.3+/-78.9]. Gamonts, which were the most common stages observed and filled the midgut and gastric caeca in grasshoppers kept in rearing rooms, had a stocky appearance and also varied greatly in size (total length: 80-348 microm; 205+/-13). Association of gamonts was precocious, biassociative, and caudofrontal. Gametocysts were spherical and highly variable in size (96-376 microm in diameter; 202.8+/-52.5), and normally have 14 sporoduct basal discs. Everted sporoducts were up to 60 microm long. Oocysts were uniformly doliform in shape, measured (5+/-0.08 by 3.2+/-0.06 microm) and contained eight sporozoites. Wall reinforcements (carinae) were present. No infection resulted in experimentally inoculated Locusta migratoria, which is a host of Gregarina acridiorum. G. ronderosi is strikingly similar to G. acridiorum, but has larger oocysts.     

Solution structure of an LNA hybridized to DNA: NMR study of the d(CT(L)GCT(L)T(L)CT(L)GC):d(GCAGAAGCAG) duplex containing four locked nucleotides

We have used two-dimensional (1)H NMR spectroscopy at 750 MHz to determine a high-resolution solution structure of an oligonucleotide containing restricted nucleotides with a 2'-O, 4'-C-methylene bridge (LNA) hybridized to the complementary DNA strand. The LNA:DNA duplex examined contained four thymidine LNA modifications (T(L), d(C1T(L)2G3C4T(L)5T(L)6C7T(L)8G9C10):d( G11C12A13G14A15A16G17C 18A19G20). A total relaxation matrix approach was used to obtain interproton distance bounds from NOESY cross-peak intensities. These distance bounds were used as restraints in molecular dynamics (rMD) calculations. Forty final structures were generated for the duplex from A-form and B-form DNA starting structures. The root-mean-square deviation (RMSD) of the coordinates for the 40 structures of the complex was 0.6 A. The sugar puckerings are averaged values of a dynamic interchange between N- and S-type conformation except in case of the locked nucleotides that were found to be fixed in the C3'-endo conformation. Among the other nucleotides in the modified strand, the furanose ring of C7 and G9 is predominantly in the N-type conformation whereas that of G3 is in a mixed conformation. The furanose rings of the nucleotides in the unmodified complementary strand are almost exclusively in the S-type conformation. Due to these different conformations of the sugars in the two strands, there is a structural strain between the A-type modified strand and the B-type unmodified complementary strand. This strain is relaxed by decreasing the value of rise and compensating with tip, buckle, and propeller twist. The values of twist vary along the strand but for a majority of the base pairs a value even lower than that of A-DNA is observed. The average twist over the sequence is 32+/-1 degrees. On the basis of the structure, we conclude that the high stability of LNA:DNA duplexes is caused by a local change of the phosphate backbone geometry that favors a higher degree of stacking.     

Occurrence of pathogens in Ips typographus (Coleoptera: Curculionidae) and in other spruce bark beetles from the wilderness reserve Dürrenstein (Lower Austria)

The study presents new data on spatial distribution of bark beetle pathogens, on changes in frequency over several years and on their prevalence during different time periods within a year from several locations within the wilderness reserve Dürrenstein (Lower Austria). The occurrence of pathogens was investigated in Ips typographus (during five years), in Pityogenes chalcographus (during two years) and in Ips amitinus (in one year). In total, seven pathogen species could be detected in I. typographus. The most dominant pathogen species were the Ips typographus-Entomopoxvirus (ItEPV), the sporozoan species Gregarina typographi and the microsporidium Chytridiopsis typographi; the latter two pathogen species were recorded every year and at about similar high (G. typographi) or low (C. typographi) rates, the ItEPV in strongly varying rates. The neogregarine Mattesia cf. schwenkei and the two microsporidia Nosema typographi and Unikaryon montanum were found in I. typographus only sporadically and the rhizopodan species Malamoeba scolyti was found once. The number of infected males and females was relatively similar with almost all pathogen species in most of the years except U. montanum, which occurred exclusively in females. Three pathogen species were recorded in P. chalcographus which were Gregarina typographi, Mattesia cf. schwenkei and Chytridiopsis typographi. Two pathogen species were observed in I. amitinus, Gregarina typographi and Chytridiopsis typographi.     

Influence of temperature on the development, reproduction and longevity of Ceratothripoides claratris (Thysanoptera: Thripidae) on tomatoes

Ceratothripoides claratris (Shumsher) is a serious pest attacking tomatoes in Thailand. Temperature-dependent development of C. claratris was studied at seven constant temperatures, i.e. 22, 25, 27, 30, 34, 35 and 40 degrees C. Pre-adult survivorship was greatest (95%) at 25 and 30 degrees C and shortest at 22 degrees C. Egg-to-adult time decreased within the range of 20 to 30 degrees C and at 34 degrees C it started to increase. The lower thermal threshold for egg-to-adult development was estimated at 16 and 18 degrees C by linear regression and the modified Logan model, respectively. The optimum temperature for egg-to-adult development was estimated at 32-33 degrees C by the modified Logan model. The influence of temperature on reproduction and longevity of C. claratris was determined at 25, 30 and 35 and 40 degrees C. Both inseminated and virgin females failed to reproduce at 40 degrees C. Virgin females produced only male offspring, confirming arrhenotoky. The sex ratio of the offspring of fertilized females was strongly female-biased, except at 25 degrees C. Mean total fecundity per female and mean daily total fecundity per female were highest for both virgin and inseminated females at 30 degrees C. Female longevity was longest at 25 degrees C and shortest at 40 degrees C. Male longevity was longest at 30 degrees C and shortest at 40 degrees C. The net reproductive rate (R0) and intrinsic rate of natural increase (rm) was greatest at 30 degrees C while, mean generation time (G) and the doubling time (t) were highest at 25 degrees C. The finite rate of increase (lambda) was fairly constant (1.1-1.5 days) over the three temperatures tested. The pest potential of C. claratris for tropical Asia is discussed.     

Biology and behavior of the seed borer wasp Bephratelloides cubensis ashmead (Hymenoptera: Eurytomidae)

The soursop Annona muricata is an important fruit for national market, and for exportation, but the crop is affected by pests and diseases. The seed borer wasp Bephratelloides cubensis Ashmead is the pest that produces the highest damage to the crop in Mexico. Sixty percent of damaged fruits and 5-50 seeds per fruit have been registered, with 25% reduction in yield. In Nayarit, Mexico, 100% of damaged fruits were recorded. In this State, an experiment with soursop was conducted to study the life cycle under field conditions and to determine diurnal behavior of the female of B. cubensis. The highest activity of the wasp was observed between 12:00h and 13:00h (35oC, 54% RH and 409.34 luxes). Females oviposited in fruits with a diameter of 3.1-7.6 cm. Larvae of B. cubensis developed five instars, adults survived no longer than 22 days, and female survived longer than males; they lived 22 and 15 days, respectively. Life cycle of B. cubensis varied from 69 to 122 days.     

黄瓜霜霉病菌侵染若干因子的研究

研究了温、湿度条件对黄瓜霜霉病菌致病性的影响,结果表明,25～35℃最适宜黄瓜霜霉病的发生,15/35℃的交替温度变化最有利于霜霉病菌的侵染,但35℃以上的高温对霜霉病菌具有杀伤作用;2 h的湿度条件就足以引起侵染,一旦侵入寄主,环境湿度条件对病害的发展影响不大.-20℃低温冷冻保存10个月和干燥放置7 d的霜霉菌种仍具致病力.发病的黄瓜叶片可以连续产生孢子囊,但随着发病时间的延长产生孢子囊量逐渐减少.活体叶片单位面积上产生的孢子囊量比离体叶片大,且显症天数与叶片单位面积产生孢囊量呈抛物线型关系.     

Effect of temperature on the development and viability of Gryon gallardoi (Brethes) (Hymenoptera: Scelionidae) parasitizing Spartocera dentiventris (Berg) (Hemiptera: Coreidae) eggs.

The development and viability of Gryon gallardoi (Brethes) (Hym.: Scelionidae) in Spartocera dentiventris (Berg) (Hem.: Coreidae) eggs were studied under four temperatures: 15, 20, 25, and 30 +/- 1 degree C, with a 12-h photophase. No parasitoid developed at 15 degrees C. Otherwise, viability reached 98.8% without varying significantly over the temperature range tested. The duration of development for males and females was inversely proportional to the temperature increase, varying respectively from 46.2 +/- 0.13 and 47.1 +/- 0.11 days (20 degrees C) to 13.3 +/- 0.07 and 13.4 +/- 0.06 days (30 degrees C). Males developed faster than females. The values estimated for the lowest thermic thresholds of development and the thermic constants were 15.5 degrees C and 185.19 DD for males and 15.6 degrees C and 192.31 DD for females, respectively. Given the average weather conditions in Porto Alegre, RS (30 degrees 01' S and 51 degrees 13' W), Brazil, G. gallardoi could annually produce 8.54 and 8.07 generations of males and females, respectively. The low rates of parasitism observed in the field during the first generation of its host may be due to the small number of G. gallardoi generations in this period.     

Thermal requirements of Galleria mellonella L. (Lepidoptera: Pyralidae) immature stages

The rearing of Galleria mellonella L. in laboratory is important for multiplication of entomopathogenic nematodes, mandatory for biological control studies. The objective of this study was to evaluate the effect of three thermal profiles on development stages of this insect, allowing synchronization of cycle production. Two distinct rearing phases were done: firstly, using nucleous of incubation for development of eggs and, secondly, using circular-aluminum manifolds for development of larvae and pupae. The time necessary for development of the immature stages decreased with higher temperatures. Incubation periods lasted 13.4 days at 22 degrees C, 8.3 at 27 degrees C and 6.8 days at 32 degrees C, while periods for larvae development lasted 40.4, 27.2, and 23.4 days, respectively, for the same temperatures. Development to pupal stage was observed 18.2, 15.0, and 12.2 days, respectively, for the same temperatures. Larval survival was higher at 32 degrees C, however embryonic stages and pupae survival were higher at 27 degrees C. and 22 degrees C, respectively. The threshold temperature was 11.209167 degrees C for the embryonic development stage, 7.695869 degrees C for larval stage, and 1.943050 degrees C for pupal stage of G. mellonella. Thermal constants were 138.380533 DG (degree day) for egg, 554.968830 DG for larvae, and 369.054080 DG for pupae.     

Some ecological parameters of Ctenocephalides felis strongylus (Jordan, 1925) (Siphonaptera: Pulicidae)

Among the fleas of medico-veterinary interest, Ctenocephalides felis (Bouché, 1 835) is the one most studied. This taxon includes two subspecies: Ctenocephalides f. felis, and Ctenocephalides f. strongylus (Jordan, 1925); only C. f. felis has been the subject of almost all the studies available. We were, thus, interested in C. f. strongylus which can be regarded as the species of substitution of C. f. felis on the African continent. The purpose of our work was to establish some biological parameters such as: hatching of eggs, cycle of development and emergence of adults. These data were compared with those available on C. f. felis. With temperatures ranging between 19 degrees C and 29 degrees C and a relative humidity (HR) of 75 % +/- 5, the hatching rates of eggs observed from the two subspecies of C. felis, are higher than 88 %. The optimal temperature of eggs hatching for C. felis is 29 degrees C, with more than 70 % of hatching obtained in 1-2 days after the laying. The larval developments of the two subspecies are almost identical and function of the temperature 18-9 days with 27 degrees C). Only differs the minimal duration of the progressive cycle. For C. f. strongylus, it lasts in 16-17 days at 29 degrees C, 20-21 days at 27 degrees C and 38 days at 19 degrees C. For C. f. felis, published values give report of 15 days at 27 degrees C and 17 days at 24 degrees C. The emergence of adults of C. f. strongylus takes eight to ten days between 19 degrees C and 29 degrees C, while data published on C. f. felis are about 26 days at 19 degrees C and 15 days at 27 degrees C.     

Biology of Agistemus brasiliensis Matioli, Ueckermann & Oliveira (Acari: Stigmaeidae) and its predation potential on Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae)

The present work aimed to study the biology of Agistemus brasiliensis Matioli, Ueckermann & Oliveira at the following temperatures, namely 15, 20, 25, 30 and 35 degrees C, fed with Brevipalpus phoenicis (Geijskes) and Typha pollen, in laboratory conditions. Life tables were calculated to evaluate the biological parameters. The optimal development of A. brasiliensis took place at 29 degrees C. The values of T (time of generation - days), R0 and r m at 30 degrees C were, 13.95, 16.25 and 0.20, respectively. The prey consumption of A. brasiliensis was studied at the densities of 5, 10, 20, 40 and 60 leprosis mite females per cage (3 cm in diameter) on citrus fruits at 29 degrees C. The maximum prey, namely 7.6 B. phoenicis females per day, were consumed at a density of 20 leprosis mites. At densities of above 40 leprosis mites per cage, A. brasiliensis oviposits 4.7 eggs per day, in comparison to 2.5 eggs per day at 20 mites per cage.     

意大利蝗和西伯利亚蝗高温耐受能力及酶活性比较研究

【目的】阐明意大利蝗Calliptamus italicus和西伯利亚蝗Gomphocerus sibiricus高温耐受能力及酶活性差异。【方法】以3℃为间隔,将意大利蝗和西伯利亚蝗成虫分别在27~48℃、24~42℃下处理4 h后测定死亡率及POD、SOD和CAT活性值。【结果】27~42℃处理下,意大利蝗存活率显著大于西伯利亚蝗(P<0.05)。雌性意大利蝗的LT50、LT90分别为48.76℃和50.67℃,雄虫则分别为47.90℃和50.53℃;雌性西伯利亚蝗的LT50、LT90分别为39.21℃和42.10℃,雄虫则分别为36.11℃和41.43℃。随着温度升高,雄性意大利蝗和雌、雄西伯利亚蝗体内POD、SOD、CAT活性先升高后降低,雌性意大利蝗体内POD活性则先降低后升高。27~42℃范围内,西伯利亚蝗体内POD和SOD活性值增幅较大,分别达到54.69%、23.54%;意大利蝗体内则以CAT活性值增幅最大,为27.02%。【结论】两种蝗虫耐高温能力存在差异,意大利蝗具有更强的耐热能力。     

Myosin-Like Protein (MR 175,000) In Gregarina Blaberae

ABSTRACT. A myosin-like protein (M_r 175,000) was detected in the parasitic protozoan Gregarina blaberae , by both immunofluorescence and immunoblotting of one- and two-dimensional electrophoresis gels using anti-myosin antibodies. This protein was present in the trophozoite ghost but not in the cytoplasmic extract, nor in extract from the sexual stage, suggesting a protein-stage-dependent expression. the protein tightly bound to the cortical membranes was insoluble at low ionic strength, or in detergent solutions, but could be extracted from Gregarina ghosts by 6 M urea in high ionic strength solution (0.5 M NaCI) and in the presence of reducing agents (20 mM DTT). the protein was localized by indirect immunofluorescence in the cortex of the epimerite, in the fibrillar disc (the socalled septum) separating the proto- and the deutomerite segments, in the contractile ring or sphincter at the top of the protomerite, and as longitudinal lines underlying the G. blaberae epicyte folds. the presence of both actin-like and myosin-like proteins would be consistent with a role in gliding and other cell motility processes of this parasite.     

Effects of Temperature and Relative Humidity on Development,Reproduction, and Predation in Feltiella acarisuga (Vallot) (Diptera: Cecidomyiidae)

The predatory gall midge Feltiella acarisuga (Vallot) (Diptera: Cecidomyiidae) is a biological control agent for twospotted spider mites on greenhouse vegetable crops. Effects of temperature and relative humidity (RH) on development of immatures, reproduction, and prey capture were determined in order to confirm the suitability of F. acarisuga for use in greenhouses. Developmental time ranged from 10 days at 27°C to 34 days at 15°C. At 20°C, developmental time was significantly shorter at 96% RH than at 84% RH. There was very poor survival of immatures at 64% RH and none at 36%. Lifespan of adult females decreased with increasing temperature, but temperature had no significant effect on number of eggs laid. At 20°C, lifespan was longer at 84 and 96% RH than at 64 or 36% RH. The number of spider mites attacked by 3-day-old larvae over 8 h increased with increasing temperature from 15 to 27°C. The number of mites attacked also increased with increasing RH at 27°C. We conclude that F. acarisuga will complete its life cycle and reproduce under conditions typically found in vegetable greenhouses in northern temperate climates. However, extended periods of low RH (<60% RH) could reduce reproduction and survivorship sufficiently to impair the predator's action against spider mite populations.