Delivery of Double-Stranded RNAs (dsRNAs) Produced by Escherichia coli HT115(DE3) for Nontransgenic RNAi-Based Insect Pest Management.

RNA interference (RNAi) is a powerful mechanism that can be exploited not only for physiology research but also for designing insect pest management approaches. Some insects cause harm by vectoring diseases dangerous to humans, livestock, or plants or by damaging crops. For at least a decade now, different insect control strategies that induce RNAi by delivering double stranded RNA (dsRNA) targeting essential genes have been proposed. Here, we focus on nontransgenic RNAi-based approaches that use oral delivery of dsRNA through feeding of inactivated bacteria to produce RNAi in disease vectors and in a crop pest. This potential pest management method could be easily adapted to target different genes or similar organisms.

A Novel Genetic Sexing Strain of Anastrepha ludens for Cost-Effective Sterile Insect Technique Applications: Improved Genetic Stability and Rearing Efficiency.

Anastrepha ludens (Loew) is one of the most destructive insect pests damaging several fruits of economic importance. The sterile insect technique (SIT) is used under an area-wide integrated pest management approach, to suppress these pest populations. Mass rearing facilities were initially established to produce sterile males of bi-sexual strains in support of SIT. The first genetic sexing strain (GSS) for A. ludens, Tapachula-7, based on pupal color dimorphism, was a key development since the release of males-only significantly increases the SIT efficiency. In this study, we document the development of a novel pupal color-based GSS. Twelve radiation-induced translocation lines were assessed as potential GSS in terms of recombination rates and rearing efficiency at a small scale. The best one, GUA10, was cytogenetically characterized: it was shown to carry a single translocation between the Y chromosome and chromosome 2, which is known to carry the black pupae marker. This GSS was further evaluated at medium and large scales regarding its genetic stability, productivity and quality versus Tapachula-7. GUA10 presented better genetic stability, fecundity, fertility, production efficiency, flying ability, and male mating, clear indicators that GUA10 GSS can significantly improve the efficacy and cost-effectiveness of SIT applications against this pest species.

The Role of Satellite DNAs in Genome Architecture and Sex Chromosome Evolution in Crambidae Moths.

Tandem repeats are important parts of eukaryotic genomes being crucial e.g., for centromere and telomere function and chromatin modulation. In Lepidoptera, knowledge of tandem repeats is very limited despite the growing number of sequenced genomes. Here we introduce seven new satellite DNAs (satDNAs), which more than doubles the number of currently known lepidopteran satDNAs. The satDNAs were identified in genomes of three species of Crambidae moths, namely Ostrinia nubilalis, Cydalima perspectalis, and Diatraea postlineella, using graph-based computational pipeline RepeatExplorer. These repeats varied in their abundance and showed high variability within and between species, although some degree of conservation was noted. The satDNAs showed a scattered distribution, often on both autosomes and sex chromosomes, with the exception of both satellites in D. postlineella, in which the satDNAs were located at a single autosomal locus. Three satDNAs were abundant on the W chromosomes of O. nubilalis and C. perspectalis, thus contributing to their differentiation from the Z chromosomes. To provide background for the in situ localization of the satDNAs, we performed a detailed cytogenetic analysis of the karyotypes of all three species. This comparative analysis revealed differences in chromosome number, number and location of rDNA clusters, and molecular differentiation of sex chromosomes.

Efectos de un programa de entrenamiento físico de 16 semanas sobre el peso, el IMC y la aptitud física en cadetes / Effects of a 16-week physical training program on weight, BMI and physical fitness in cadets / Efeitos de um programa de treinamento físico de 16 semanas sobre peso, IMC e condicionamento físico em cadetes

OBJETIVO: Evaluar los efectos de un programa de entrenamiento físico de 16 semanas sobre el peso, el IMC y la aptitud física en cadetes. MÉTODO: Cincuenta cadetes pertenecientes al Instituto Estatal de Ciencias Penales y Seguridad Pública del Estado de Sinaloa (México) fueron divididos en dos grupos: grupo de ejercicio (n = 36, media de 26.1 años) y grupo control (n = 14 media, de 26.7 años). Se determinaron el peso y el Índice de Masa Corporal como medida corporal y el Consumo Máximo de Oxígeno, fuerza abdominal, flexo-extensión del codo y velocidad en 40m. para la aptitud física. Se realizaron evaluaciones antes y después de un programa de entrenamiento físico de 16 semanas. RESULTADOS: Los cadetes presentaron aceptables ganancias en el Consumo Máximo de Oxígeno, fuerza abdominal, fuerza flexo-extensora del codo y velocidad de carrera tras el entrenamiento, pero el peso y el Índice de Masa Corporal pesar que se obtuvieron mejoras tras el periodo de entrenamiento estas mejoras no difieren significativamente del grupo control. El grupo control no mostró cambios en ninguna variable estudiada. CONCLUSIONES: Un programa de entrenamiento físico de 16 semanas mejora todos los componentes de la aptitud física, pero el peso y el Índice de Masa Corporal presentaron ligeras modificaciones

OBJECTIVE: To assess the effects of a 16-week physical training program on weight, Body Mass Index and physical fitness in cadets. METHOD: Fifty cadets belonging to the State Institute of Criminal Sciences and Public Security of the State of Sinaloa (Mexico) divided into two groups: exercise group (n = 36, mean of 26.1 years) and control group (n = 14, mean of 26.7 years). Weight and Body Mass Index were measured as body measurements and Maximal Oxygen Uptake, abdominal strength, flexion-extension of the elbow and speed in 40m. for physical fitness. Evaluations were conducted before and after a 16-week physical training program. RESULTS: The cadets had acceptable gains in Maximal Oxygen Uptake, abdominal strength, flexion-extensor elbow strength and running speed after training, but the weight and the Body Mass Index, although improvements were obtained after the training period, these improvements do not differ significantly from the control group. The control group did not show changes in any variable studied. CONCLUSIONS: A 16-week physical training program improved all components of physical fitness, but the weight and the Body Mass Index showed slight modifications

OBJETIVO: Avaliar os efeitos de um programa de treinamento físico de 16 semanas sobre peso, Índice de Massa Corporal e aptidão física em cadetes. MÉTODO: Cinquenta cadetes pertencentes ao Instituto Estadual de Ciências Criminais e Segurança Pública do Estado de Sinaloa (México) divididos em dois grupos: grupo de exercícios (n = 36, média de 26.1 anos) e grupo de controle (n = 14, média de 26.7 anos). Peso e Índice de Massa Corporal foram medidos como medidas corporais e Máxima Absorção de Oxigênio, força abdominal, flexo-extensão do cotovelo e velocidade em 40m. para aptidão física. As avaliações foram realizadas antes e depois de um programa de treinamento físico de 16 semanas. RESULTADOS: Os cadetes tiveram ganhos aceitáveis de Máxima Absorção de Oxigênio, força abdominal, força de flexão-extensor do cotovelo e velocidade de corrida, mas pequenas melhorias no peso e no Índice de Massa Corporal após o programa de treinamento após o treinamento, mas o peso e o Índice de Massa Corporal, embora as melhorias tenham sido obtidas após o período de treinamento, essas melhorias não diferem significativamente do grupo controle. CONCLUSÕES: Um programa de treinamento físico de 16 semanas melhorou todos os componentes da aptidão física, mas o peso e o Índice de Massa Corporal mostraram pequenas modificações

Evaluation of horizontal gene transfer risk between the Mediterranean fruit fly Ceratitis capitata (Tephritidae) and its parasitoid Fopius ceratitivorus (Braconidae).

The transgenic strain of the Mediterranean fruit fly (medfly), Ceratitis capitata (Wied.) VIENNA 8 1260, developed from the classical genetic sexing strain VIENNA 8, has two molecular markers that exhibit red fluorescence in the body and green fluorescence in testicles and sperm. These traits offer a precise tool to discriminate between mass-reared sterile males and wild fertile males, and they could potentially increase the effectiveness of control programs for this pest. To assess the risk of horizontal transfer of the fluorescence transgenes in natural ecosystems, we used the VIENNA 8 1260 strain and the medfly parasitoid Fopius ceratitivorus. The fluorescence signal and the inheritance of the fluorescence gene markers were monitored for over 16 generations (about two years) in both species using fluorescence microscopy and a PCR-based assay. The PCR analysis was performed in four independent laboratories. Both fluorescence microscopy and PCR analysis indicated that no horizontal gene transfer of the DsRed transgene occurred during 16 generations of medfly parasitoid rearing under experimental conditions.

Suppression of Mediterranean Fruit Fly (Diptera: Tephritidae) With Trimedlure and Biolure Dispensers in Coffea arabica (Gentianales: Rubiaceae) in Hawaii.

To assess the potential to suppress Mediterranean fruit fly, Ceratitis capitata (Wiedemann; Diptera: Tephritidae), via mass trapping with Trimedlure (TML), we compared fly catch (as catch per trap per time period) provided by either a novel, solid, triple-lure dispenser with TML, methyl eugenol (ME), and raspberry ketone (RK) (TMR) or solid TML plugs, both without insecticides, in addition to Biolure bait stations. Work was done in a coffee plantation that had a dense C. capitata population. Three treatments were compared: 1) TMR or TML (50 traps per ha), 2) Biolure (50 traps per ha), 3) TML (25 per ha) or TMR (25 per ha) + Biolure (25 per ha), and 4) an untreated control. During coffee season, based on C. capitata captures (mean flies per trap per wk) inside plastic McPhail traps, all treatments were significantly different than the control: Biolure (9.57) = TMR (11.28) = Biolure +TMR (13.50) < Control (36.06 flies/trap/wk). During non-coffee season, all treatments were significantly different than the control and TML was significantly lower than Biolure (wax matrix bait stations): TML (0.95) < Biolure (1.43) = Biolure +TML (1.77) < Control (2.81 flies/trap/wk). Surprisingly, captures were not lower in plots treated with combinations of Biolure + TMR or TML, compared to individual plots with Biolure or TML or TMR alone. Mass trapping with either TML or TMR dispensers deserves further study as a component of Integrated Pest Management programs for C. capitata in Hawaii and may have global potential for management of C. capitata.

Effect of Irradiation Doses on Sterility and Biological Security in a Genetically Modified Strain of the Mediterranean Fruit Fly (Diptera: Tephritidae).

The genetically modified strain of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) VIENNA 8 1260, was developed from the genetic sexing strain VIENNA 8. It has two molecular markers that exhibit red fluorescence in the body and green fluorescence in testis and sperm. These traits offer a precise tool to discriminate between mass-reared and wild males, increasing the effectiveness of sterile insect technique. The reproductive performance of the VIENNA 8 1260 and VIENNA 8 D53- (without the D53 inversion introduced to prevent recombination) was compared at different irradiation doses. The general effect of irradiation on VIENNA 8 1260 followed the same patterns documented in previous publications for VIENNA 8 D53-. Irradiation doses of 80 Gray or greater reduced fertility and induced high levels of sterility in wild females. Fecundity reduction was higher in VIENNA 8 1260 than in VIENNA 8 D53- females. Vertical transmission of the fluorescence gene was confirmed up to the F4 generation. Substerilization in the VIENNA 8 1260 could jeopardize the usefulness of the transgenic strain due to the possible vertical transfer of the fluorescence transgene from the sterile males to the wild flies. A biologically safe higher irradiation dose could result in reduced competitiveness of the VIENNA 8 1260 strain. Mating and remating experiments suggest that Mediterranean fruit fly females exhibit a relative precedence in the use of the sperm: though both sperms are mixed, sperm from the remating is spent first. Results suggest a lower fitness of VIENNA 8 1260 sperm, when compared with sperm from a nonfluorescent bisexual strain, which is consistent with the lower reproductive performance documented for the VIENNA 8 1260 strain.

Performance of a Genetically Modified Strain of the Mediterranean Fruit Fly (Diptera: Tephritidae) for Area-Wide Integrated Pest Management With the Sterile Insect Technique.

The genetically modified strain of Ceratitis capitata (Wiedemann) VIENNA 8 1260 has two morphological markers that exhibit fluorescence in body and sperm. To assess the feasibility of its use in area-wide integrated pest management (AW-IPM) programs using the sterile insect technique, its rearing performance and quality control profile under small, medium, and large scales was evaluated, as well as in field cages. The VIENNA 8 1260 strain had a lower yield than the control strains, VIENNA 8 with D53 inversion (VIENNA 8) and without D53 inversion (VIENNA 8 D53-). At mass-rearing scale, yield gradually increased in three generations without reaching the control strain values. The VIENNA 8 1260 strain was stable in the genetic sexing mechanism (>99.9%) and expression of fluorescence (100%). In field cages, the VIENNA 8 1260 males reduced the mating potential of wild males in the same magnitude as the VIENNA 8, when evaluated in independent cage tests. However, the relative sterility index and the strain male relative performance index of VIENNA 8 1260 males were significantly lower than those of the VIENNA 8. There were no significant differences in longevity of these strains. The potential application of the VIENNA 8 1260 in AW-IPM programs is further discussed.

Performance of Psyttalia humilis (Hymenoptera: Braconidae) reared from irradiated host on olive fruit fly (Diptera: Tephritidae) in California.

The parasitoid Psyttalia humilis (Silvestri) was reared on Mediterranean fruit fly, Ceratitis capitata (Wiedemann), third instars irradiated at 0-70 Gy at the USDA, APHIS, PPQ, Moscamed biological control laboratory in San Miguel Petapa, Guatemala, and shipped to the USDA, ARS, Parlier, CA. Irradiation dose did not affect the parasitoid's offspring sex ratio (53-62% females), percentage of unemerged adults (12-34%), number of progeny produced per female (1.4-1.8), and parasitism (19-24%). Host irradiation dose had no significant effect on the forewing length of female P. humilis and its parasitism on olive fruit fly, Bactrocera oleae (Rossi) and offspring sex ratio, but dissection of 1-wk-old female parasitoids reared from hosts irradiated with 70 Gy had a significantly lower number of mature eggs than females from nonirradiated hosts. Longevity of P. humilis adults decreased with increased temperature from 15 to 35°C, regardless of food provisions, gender, and host irradiation dose. Females survived 37-49 d at 15°C with water and food, and only 1-2 d at 35°C without food, whereas males lived shorter than females at all temperatures and food combinations tested. Adult P. humilis reared from fertile C. capitata and aspirated for dispensing in cups lived significantly longer after shipment than those specimens chilled and dispensed by weight. At 21 and 32°C, 50% of parasitoids departed release cages after 180 and 30 min, respectively, but none departed at 12°C. Thirteen shipments of P. humilis (2,980-21,922 parasitoids per shipment) were received between September and December 2009, and seven shipments (7,502-22,560 parasitoids per shipment) were received between October and December 2010 from San Miguel Petapa, Guatemala. Daily number of olive fruit fly adult and percentage female trap captures ranged <1-19 and 8-58% in 2009, and <1-11 and 0-42% in 2010, respectively. The number of parasitoids released ranged 848-12,257 in 2009 and 3,675-11,154 in 2010. Percentage parasitism of olive fruit fly third instars at all locations ranged 0-9% in 2009 and 0-36% in 2010.

Efficacy of wax matrix bait stations for Mediterranean fruit flies (Diptera: Tephritidae).

Tests were conducted that evaluated efficacy of wax matrix bait stations for Ceratitis capitata (Wiedemann) adults in Guatemala. Bait stations were exposed to outdoor conditions to determine effect of weathering on longevity as indicated by bait station age. Results of laboratory tests found that bait stations with spinosad and ammonium acetate remained effective for at least 31 d compared with pesticide-free controls, although there was some loss of efficacy over time. Percentage mortality for bait station strips with 2% spinosad and 1% ammonium acetate decreased from 100 +/- 0.0% on day 0 to 70 +/- 7.1% after 31 d. Ammonia concentration had little effect on percentage mortality although there was some indication that ammonia concentration affected number of flies observed on the bait stations. Bait station strips (one per cage) were more effective than controls for 6-8 wk when tests were conducted in field cages (3 m diameter x 2 m), but only 2-3 wk when tests were conducted in large (2.5 m high and 6.0 m wide and 7.5 m long) field cages. Longevity was restored when multiple bait stations (3, 6, or 12) were deployed per cage. Bait stations containing methomyl were used for field tests of efficacy for wild flies. Dipped lure bait stations, which were made by coating two edges of commercial ammonium acetate and trimethylamine lures, killed six times more flies than corn cob bait stations dipped into a Nulure/malathion solution. They also killed more flies than pesticide-free controls for 8 wk.

Psyttalia cf. concolor (Hymenoptera: Braconidae) for biological control of olive fruit fly (Diptera: Tephritidae) in California.

The larval parasitoid, Psyttalia cf. concolor (Szépligeti), reared on Mediterranean fruit fly, Ceratitis capitata (Weidemann), by the USDA-APHIS-PPQ, Guatemala City, Guatemala, was imported into California for biological control of olive fruit fly, Bactrocera oleae (Gmelin), in olives, Olea europaea L. Mean percentage parasitism of olive fruit fly third instars infesting fruit in field cages ranged from 7.0 in Grapevine to 59.7 in Santa Barbara and in free releases ranged from 0 in Grapevine to 10.6 in Santa Barbara after 4- to 6-d exposures. In the laboratory, more parasitoids developed to adults in olive fruit fly larvae that were 11-13 d old than in larvae 8-10 d old. Adult parasitoids lived significantly longer when provided with water than adults without water in environmental chambers at 5 degrees C, 85% RH; 15 degrees C, 65% RH; 25 degrees C, 25% RH; and 35 degrees C, 25% RH. Adult parasitoids lived for 48 d with honey for food and water and 32 d with food and sugar solution at 15 degrees C and 65% RH. Survival of adult parasitoids without food and water in greenhouse tests was approximately 4 d in a simulated coastal climate and 1 d in a simulated inland valley climate and was significantly increased by providing food and water. The parasitoid did not develop in the beneficial seedhead fly, Chaetorellia succinea (Costa), in yellow star thistle. The rate of parasitism of walnut husk fly, Rhagoletis completa Cresson, larvae in green walnut husks was 28.4% in laboratory no-choice tests. In choice tests, the rate of parasitism of walnut husk fly versus olive fruit fly larvae in olives was 11.5 and 24.2%, respectively.

Capture of Mediterranean fruit flies (Diptera: Tephritidae) in dry traps baited with a food-based attractant and Jackson traps baited with trimedlure during sterile male release in Guatemala.

Captures of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann), in Jackson traps baited with trimedlure were compared with captures in cylindrical open-bottom dry traps baited with a food-based synthetic attractant (ammonium acetate, putrescine, and trimethylamine). Tests were conducted in Guatemala during a sterile male release program in an area where wild flies were present in low numbers. More wild and sterile females were captured in food-based traps, and more wild and sterile males were captured in trimedlure traps. The food-based traps captured almost twice as many total (male plus female) wild flies as the trimedlure traps, but the difference was not significant. Females made up approximately 60% of the wild flies caught in the food-based attractant traps; the trimedlure traps caught no females. The ratio of capture of males in trimedlure traps to food-based traps was 6.5:1 for sterile and 1.7:1 for wild flies. Because fewer sterile males are captured in the food-based traps, there is a reduction in the labor-intensive process of examining flies for sterility. The results indicate that traps baited with food-based attractants could be used in place of the Jackson/trimedlure traps for C. capitata sterile release programs because they can monitor distributions of sterile releases and detect wild fly populations effectively; both critical components of fruit fly eradication programs by using the sterile insect technique.

Effects of diet, ginger root oil, and elevation on the mating competitiveness of male Mediterranean fruit flies (Diptera: Tephritidae) from a mass-reared, genetic sexing strain in Guatemala.

The release of sterile males is a key component of an areawide program to eradicate the Mediterranean fruit fly, Ceratitis capitata (Wiedemann), from Guatemala and southern Mexico. The objective of our study was to assess the effects of adult diet, exposure to ginger root oil (Zingiber officinale Roscoe), and elevation on the mating competitiveness of the sterile males used in an areawide program. Sterile males were maintained on a protein-sugar (protein-fed) or a sugar-only (protein-deprived) diet and were exposed (for 4 h 1 d before testing) or not exposed to ginger root oil. In field-cage trials conducted at a high (1,500 m) and low (700 m) site, we monitored the influence of these treatments on the mating success of sterile males in competition with wild males (reared exclusively on the protein-sugar diet and without ginger root oil exposure) for wild females. Elevation and ginger root oil exposure had significant effects, with sterile males having higher mating success at the low-elevation site and ginger root oil-exposed males having greater success than ginger root oil-deprived males at both sites. Diet did not have a significant overall effect, and its influence varied with elevation (dietary protein seemed to provide an advantage at the high-elevation site but not at the low-elevation site). Possible implications of these findings for eradication programs against the Mediterranean fruit fly are discussed.

Coartación de la arteria pulmonar y prolapso valvular mitral: presentación de un caso / Pulmonary artery coartation and prolapse of mitral valve: report of a case

Se presenta el caso de una paciente de 33 años de edad con coartación de la arteria pulmonar que se acompañó de prolapso de válvula mitral, y con cuadro clínico de hipertensión pulmonar. Se realizaron estudios inmunológicos y aortograma para descartar arteritis. La coartación de la arteria pulmonar es una enfermedad rara, y no se había informado previamente su ocurrencia simultánea con prolapso mitral