Insect intestinal mucins (McIIM2-4) expressed in the midgut of feeding, starved and moulting Mamestra configurata larvae were identified. McIIM2 and McIIM4 were associated with the peritrophic matrix (PM). PMs from feeding and starved larvae were translucent and contained organized chitin bundles perpendicular to their long axis, whereas PM from moulting larvae consisted of an inner opaque mass surrounded by an outer translucent sleeve. Serine protease genes (McSP1, McSP2, McSP25 and McSP29) were also expressed in these larvae and several serine proteases were associated with the PM. Serine protease activity was also detected in the midgut of feeding, starved and moulting larvae.

Lygus Hahn plant bugs (Hemiptera: Miridae) are serious pests of a wide variety of economically important crops in North America. European Peristenus digoneutis Loan and P. relictus Ruthe (Hymenoptera: Braconidae) are being considered for release in Canada as part of a classical biological control program for Lygus. The attractiveness of different host plants to European Peristenus has not been addressed, but may be an important consideration prior to parasitoid release. Lygus rugulipennis Poppius nymphs were collected in the Northern Temperate Atlantic (NTA) ecoregion on red clover (Trifolium pratense L.; Fabaceae) and chamomile (Matricaria recutita L.; Asteraceae), and in the Western European Broadleaf Forest (WEBF) ecoregion on red clover and alfalfa (Medicago sativa L.; Fabaceae). Parasitism levels and parasitoid species were determined using a multiplex PCR assay for P. digoneutis, P. relictus, and P. pallipes Curtis. Mean parasitism levels in L. rugulipennis were 45-49% in the NTA ecoregion and 25-32% in the WEBF ecoregion. However, in neither ecoregion were parasitism levels and parasitoid species compositions significantly different in nymphs from different host plant species. Furthermore, multiparasitism was low despite the fact that P. digoneutis and P. relictus share the same host species.

One- and two-dimensional gel electrophoresis coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to identify cDNA encoding a chitin deacetylase (McCDA1) and three insect intestinal lipases (McIIL1, McIIL2 and McIIL3) associated with the Mamestra configurata (bertha armyworm) peritrophic matrix. Recombinant McCDA1 was active and chitin deacetylase activities were detected in the midgut. McCDA1 and the McIIL genes were expressed exclusively in the midgut; however, McCDA1 and McIIL2 were expressed in all larval stages, whereas McIIL1 was expressed mainly in feeding larvae and McIIL3 primarily during the moult.

Lygus spp. Hahn (Hemiptera: Miridae) are serious pests of a wide variety of economically important crops in North America. A European parasitoid, Peristenus digoneutis Loan (Hymenoptera: Braconidae), released in the northeastern USA for the biological control of Lygus, has successfully established in nine states and in eastern Canada, including southeastern Ontario, southern Quebec and Nova Scotia. To determine the extent to which P. digoneutis has dispersed and established in mirid populations in Ontario, a single-step multiplex PCR assay, designed to differentiate European species P. digoneutis and P. relictus from native North American Peristenus species, was used to identify parasitoid larvae dissected from field-collected Miridae. A total of 222 parasitoid larvae were analysed with the multiplex assay. Most (172) were identified as members of the P. pallipes Curtis complex; however, three from the Niagara region were identified as P. digoneutis. Specimens that did not amplify using the Peristenus multiplex PCR assay were screened with a PCR primer set designed to detect Leiophron (Hymenoptera: Braconidae) species, and most were confirmed to be Leiophron larvae. This study confirms the presence of P. digoneutis species in southern Ontario, and is another example of the utility of molecular methods for the detection of newly introduced or dispersed parasitoids. The presence of exotic P. digoneutis in southern Ontario may expedite future releases to augment already-established populations. Following such releases, the Peristenus multiplex PCR assay will be a useful component of post-release studies to evaluate the success of the biological control programme for Lygus plant bugs in Canada.

The polymerase chain reaction (PCR) revolutionized the field of diagnostics, and today it has routine applications in medical, veterinary, forensic and botanical sciences. The fields of biological control and insect pest management have generally been slow to adopt PCR-based diagnostics in comparison with other fields of science. However, there has been increasing interest in the use of molecular diagnostic tools in arthropod biological control. In applied entomology, molecular techniques have generally been used for insect identification and systematics; however, PCR-based techniques are increasingly becoming recognized as valuable tools in ecological studies. Here, we review research that has used PCR-based techniques for parasitoid and predator/prey identification and detection, and place these studies in the context of their contributions to biological control of arthropods. The status and future directions of diagnostic molecular markers in applied entomology and insect pest management are also discussed.

Laboratory experiments were conducted to evaluate the contact and oral toxicity of commercial formulations of spinosad and deltamethrin to adults of the crucifer flea beetle, Phyllotreta cruciferae (Goeze). Method of exposure had a significant effect on flea beetle mortality and feeding damage to canola seedlings. Topical treatment of flea beetles with deltamethrin or different concentrations of spinosad resulted in significantly lower mortality and higher feeding damage than exposure to treated canola cotyledons. Results indicated that spinosad was more toxic by ingestion than by topical contact. Mortality from treated cotyledons was significantly higher with 60 ppm deltamethrin than with 80 or 120 ppm spinosad after 24 h exposure but not after 120 h exposure. Delayed mortality in the spinosad treatments did not result in high feeding damage; damage after 120 h was not significantly different in the spinosad and deltamethrin treatments. Low concentrations of spinosad (40 ppm) strongly inhibited feeding activity within 24 h after exposure. Mortality from spinosad was higher after beetles were exposed to treated cotyledons for 120 h than for 24 h. Mortality from spinosad, but not deltamethrin, was significantly higher at 25 °C than at 15 °C. An ionic surfactant, polyethylenimine, increased the toxicity of 40 ppm spinosad. Our study suggests that spinosad has potential for use as an insecticide against crucifer flea beetles on canola.

Lygus Hahn ( Hemiptera: Miridae) are serious pests of agricultural and greenhouse crops throughout North America. In Europe, bivoltine Peristenus Forster ( Hymenoptera: Braconidae) species have a significant impact on Lygus populations. Release and establishment of European P. digoneutis Loan in Lygus lineolaris ( Palisot de Beauvois) populations in northeastern USA has renewed interest in the intended liberation of European parasitoids for Lygus control in Canada. Accurate identification of natural enemies is the cornerstone of biological control but conventional methods for identifying Peristenus species and estimating parasitism rates rely on tedious and time- consuming dissection and rearing methods. The present study describes species- specific PCR primers for three species of Peristenus, and the use of a multiplex PCR assay to detect P. digoneutis and P. stygicus Loan eggs and larvae from Lygus rugulipennis Poppius nymphs. Results indicate that the primers amplify uniquely sized, species- specific PCR products for the three species and are capable of detecting single eggs in parasitized nymphs within 3 days post- parasitism. Using a multiplex PCR assay, the primers maintain specificity and sensitivity, and allow detection of each of the three species in a single reaction. Although molecular diagnostics have previously been used in the identification of parasitoids and estimation of parasitism rates, this is the first time a single- step multiplex PCR protocol has been described.

The oviposition biology of the bertha armyworm, Mamestra configurata Walker, was studied with emphasis on the effect of conspecific eggs on oviposition site selection. Bertha armyworm lay clusters of up to 700 eggs, and larvae have feeding and growth habits similar to those of other Lepidoptera that gain advantages from feeding aggregations. In a field-cage experiment, multiple egg masses per leaf were noted, although the vast majority (85%+) of leaves available for oviposition received no eggs. A series of dual-choice laboratory experiments was conducted using paired excised leaves with and without eggs or egg-wash extracts. Females strongly preferred to oviposit on leaves with eggs of a different female than on leaves without eggs. However, females did not prefer leaves with their own eggs over control leaves without eggs. Gravid females also preferred leaves treated with a methanol egg-wash over leaves treated only with methanol, indicating that the source of oviposition stimulation may be chemically based. The potential relevance of these observations is discussed in the context of host-plant distribution and their exploitation by bertha armyworm.

The peritrophic matrix (PM) of Plutella xylostelia larvae was found to contain twelve integral and eighteen loosely associated proteins. An antiserum against Mamestra configurata integral PM proteins cross-reacted with several P xylostelia PM proteins and was used to isolate a partial cDNA encoding an insect intestinal mucin (PxIIM). PxIIM was expressed primarily in the larval midgut. The deduced protein sequence of the partial cDNA contained three potentially glycosylated, mucin-like domains and six cysteine-rich chitin-binding domains (CBDs). An additional chitin-binding domain was proposed to reside at the amino terminus of the protein based on comparison with other IIM. The organization of mucin domains and CBDs exhibited features, including an internal triplet of regularly spaced CBDs and a carboxyl terminal CBD with two additional conserved cysteine residues, that were found to be common to other lepidopteran IIMs.

Infertility affects 13-18% of couples and growing evidence from clinical and epidemiological studies suggests an increasing incidence of male reproductive problems. The pathogenesis of male infertility can be reflected by defective spermatogenesis due to pituitary disorders, testicular cancer, germ cell aplasia, varicocele and environmental factors or to defective sperm transport due to congenital abnormalities or immunological and neurogenic factors. Recent studies suggest an increased incidence of genetic disorders related to male infertility which may affect different levels, interfering with germ cell generation and maturation or leading to the production of non-functional spermatozoa. The identification of genetic causes of male infertility raises the issue of the transmission of defects to the offspring, a situation that is becoming more important given the increasing use of intracytoplasmic sperm injection (ICSI), a procedure in which the natural selection of the spermatozoa is by-passed. Fertilization can occur in vitro using ejaculated, epididymal or testicular spermatozoa, either fresh or frozen-thawed, providing opportunities hitherto not possible for men to be genetic fathers.

Objective: To assess whether women can be assigned to intrauterine insemination (IUI) treatment on the basis of fallopian tubal patency, diagnosed by Hysterosalpingo Contrast Sonography (Hy Co Sy).Design: Case controlled, clinical study.Setting: Tertiary referral center.Patient(s): The cases were consecutive, infertile women who underwent Hy Co Sy and IUI. The control group was women who had IUI over the same period but whose tubal patency was assessed by hysterosalpingogram (HSG) or laparoscopy and dye (lap and dye).Intervention(s): Women with bilateral patency at Hy Co Sy and with unexplained, anovulatory, or male factor infertility underwent IUI using the partner's (IUI-H) or donor's (IUI-D) semen. Their outcome was compared with that of cohorts of women who had been examined using HSG or lap and dye.Main Outcome Measure(s): Clinical pregnancy rate per cycle and cumulative pregnancy rates at IUI-H or IUI-D.Result(s): The clinical pregnancy rates per cycle at IUI-H or IUI-D did not differ among the three groups. The cumulative pregnancy rates after three cycles of IUI-H were 0.17, 0.15, and 0.17 in the Hy Co Sy, HSG, and lap and dye cohorts, respectively, and 0.69, 0.77, and 0.54 in the same groups after six cycles of IUI-D. There were no differences in the clinical characteristics or stimulation regimes used.Conclusion(s): Women screened as "normal" by Hy Co Sy may be allocated to treatments that rely on accurate assessment of tubal patency without compromising their chance of conception. (C)2003 by American Society for Reproductive Medicine.

Commercial farms were examined in western Canada to assess the impact of two cropping systems (annual-grain and diversified grain-forage) and two input systems (high level and organic) on carabid beetles.

BACKGROUND: Withholding gonadotrophins (coasting) can reduce the risk of severe ovarian hyperstimulation syndrome (OHSS) in patients having assisted reproduction therapy. This requires daily serum estradiol (E-2) measurements, which occasionally have been seen to decline suddenly and sharply. METHODS: To increase the sensitivity of the coasting programme we measured serum FSH in parallel with E-2 in patients at risk of developing OHSS. RESULTS: Out of a total of 1240 cycles, 106 were coasted and in 89 both serum E-2 and FSH were measured at least twice during the coasting period. One case of late severe OHSS was encountered in the study group. The serum FSH declined by a rate of 24.3+/-4.5% per day. Serum E-2 level reached a 'safe level' of <10 000 pmol/l when the serum FSH declined to 5 IU/l or less. CONCLUSION: The results from this study show that measuring serum E-2 and FSH can assist in predicting the point at which serum E-2 has declined to a level safe enough to administer the trigger HCG.

Secretions of male accessory glands contain a variety of bioactive molecules. When transferred during mating, these molecules exert wide-ranging effects on female reproductive activity and they improve the male's chances of siring a significant proportion of the female's offspring. The accessory gland secretions may affect virtually all aspects of the female's reproductive activity. The secretions may render her unwilling or unable to remate for some time, facilitating sperm storage and ensuring that any eggs laid will be fertilized by that male's sperm. They may stimulate an increase in the number and rate of development of eggs and modulate ovulation and/or oviposition. Antimicrobial agents in the secretions ensure that the female reproductive tract is a hospitable environment during sperm transfer. In a few species the secretions include noxious chemicals. These are sequestered by developing eggs that are thereby protected from predators and pathogens when laid.

Host plant resistance research to date indicates that Brassica plants expressing the glossy leaf wax characteristic show some resistance to diamondback moth, Plutella zylostella (L.). In the present study oviposition and first-instar feeding preferences were examined on ‘Glossy’ and ‘Waxy’ near isogenic lines of Brassica rapa (L.). Feeding preference, growth and survival of later instar diamondback moth larvae also were examined. Although females did not discriminate between ‘Waxy’ and ‘Glossy’ plants for oviposition, there was a strong preference among first-instar larvae for ‘Waxy’ plants in the choice experiment. There were no significant differences in larval survival or fourth-instar feeding preference on any of the lines tested. The present experiment indicates that B. rapa expressing the glossy leaf wax characteristic shows some resistance to diamondback moth, similar to that observed previously with glossy B. oleracea. The resistance appears to have a behavioural basis and is expressed against early instar larvae.

The efficacy and safety of short acting buserelin and nafarelin intranasal spray were compared to long acting leuprorelin depot intramuscular or subcutaneous injection in this prospective study of 157 women undergoing controlled ovarian hyperstimulation (COH) for in-vitro fertilisation (IVF). Patients were allocated to three groups to receive buserelin 150 microg nasal spray three times daily (Group B), nafarelin nasal spray 400 microg twice daily (Group N), or leuprorelin depot 3.75 mg once by intramuscular or subcutaneous injection (Group L) for pituitary desensitisation prior to commencing COH with human menopausal gonadotrophins (hMG) according to the Centre's protocol. The mean (+/-S.D.) age (years) (32.6+/-3.8: Group B, 32.1+/-3.3: Group N versus 32.1+/-3.3: Group L); mean (+/-S.D.) total dosage of hMG (ampoules) (37.5+/-16.1: Group B, 39.8+/-14.2: Group N versus 41.9+/-12.6: Group L) and mean daily dosage of hMG (ampoules) (3.1: Group B, 2.8: Group N versus 3.0: Group L) seen were not statistically significantly different. The duration between starting the different gonadotrophin-releasing hormone (GnRHa) and the beginning of the next menstrual period was also not seen to be statistically significantly different between the three groups (Group B: 10+/-5.5, Group N: 9.1+/-4.1 versus Group L: 8.2+/-3, days). The number of abandoned cycles was higher in Group L (17% versus 11.8%: Group B and 11.3%: Group N) but this difference did not reach statistical significance. The clinical pregnancy rates per oocyte retrieval and per embryo transfer procedure were respectively, 31.1, 35% in Group B, 12.8, 14% in Group N versus 20.5, 23.7 in Group L and were not seen to be statistically significantly different even when ongoing pregnancy rates were compared. Apart from a statistically significantly greater incidence of allergic nasal reactions in the nafarelin group (P=0.001), all other side-effects were not shown to be statistically significantly different between the three groups. We conclude that a single dose of leuprorelin depot can be considered to be as an equally effective alternative to multiple doses of buserelin or nafarelin for pituitary desensitisation in women undergoing COH for IVF.

The concentration of ferulic acid (FA), the major phenolic acid in the wheat kernel, was found to differ significantly in the mature grain of six wheat cultivars known to have a range of tolerance to the orange wheat blossom midge (Sitodiplosis mosellana). Differences in FA content were correlated with floret infestation level of the cultivars. The wheat cultivars ranked similarly in FA content at the four locations where they were tested, despite a significant effect of environment. Ferulic acid was synthesized mainly during the early stages of grain filling but at different rates among cultivars. Ferulic acid was concentrated primarily in the shorts and bran fractions in an insoluble-bound form. A high correlation was obtained between FA contents as determined by GLC, fluorometry, UV, and colorimetry. The colorimetric procedure was modified as a qualitative, simple, and rapid test for identifying midge-resistant wheat and evaluated in several field trials. The method should provide a rapid tool in the preliminary screening of experimental lines in the development of midge-resistant wheat cultivars.

The growth (increase in body mass) and development (progression to specific instar) of the bertha armyworm, Mamestra configurata Walker, were examined using up to 14 plant lines from five species of brassicaceous plants. Varietal and breeding lines of Brassica napus L., B. juncea L., B. I;apa L., B. catinata L., and Sinapis alba L. were selected on the basis of differing seed and foliar glucosinolate levels. No-choice feeding experiments were conducted with larvae isolated on leaf discs, individual leaves, or entire plants of each line. The no-choice tests showed that M. configuruta weight gain and development varied significantly among host plants tested. Two B. juncea lines (AC Vulcan and H-Allyl) and the S. alba lines (AC Pennant and L-GS) were the poorest, hosts in terms of larval weight grain, and the two B. juncea lines also significantly slowed development relative to the other lines tested. Mamestra configurata feeding preference, which was examined using a series of paired leaf disc choice tests, was also significantly different among the lines examined. Two B. juncea lines (AC Vulcan and H-Allyl) were signifcantly less preferred than all the other lines examined. The data extends previous work indicating that specific foliar glucosinolates, such as sinigrin, which is predominant in B. juncea, and sinalbin, which is abundant in S. alba, may provide brassicaceous crops with some protection from M. configurata. Thus, it may be possible to use foliar glucosinolate levels as predictors of M. configurata feeding damage in the development of breeding lines for the genus Brassica.

The isolated oviduct of the migratory grasshopper, Melanoplus sanguinipes, shows a myogenic, rhythmic pattern of contraction. The pattern of contraction can be modified by treatment with hemolymph or extracts from a variety of tissues from animals of differing age, sex, and mated status. Extracts of oviducts and ventral nerve cord (VNC), as well as hemolymph, from both virgin and mated females were almost always stimulating, though their effect on frequency, amplitude and/or tonus varied. In contrast, whereas extracts of spermatheca and brain from mated females were stimulating, extracts of these tissues from virgin females inhibited contraction. All male material tested (long hyaline tubules (LHT), the male accessory gland complex less the LHT, testes, brain, VNC and hemolymph) stimulated oviduct contraction in a dose-dependent manner, usually enhancing at least two of frequency, amplitude and tonus. However, oviposition-stimulating protein, a purified product of the LHT, provoked only an increase in frequency when applied to the oviduct. LHT extract modulated the effects of virgin female tissue extracts, always in a stimulatory manner.