Mass-Rearing Conditions Do Not Always Reduce Genetic Diversity: The Case of the Mexican Fruit Fly, Anastrepha ludens (Diptera: Tephritidae).

The application of the sterile insect technique (SIT) requires the adaptation of insects to mass-rearing conditions. It is generally accepted that this adaptation may include a reduction in genetic diversity and an associated loss of desirable characteristics for the effective performance of sterile insects in the field. Here, we compare the genetic diversity of two mass-reared strains of the Mexican fruit fly, Anastrepha ludens, and a wild (WIL) population collected near Tapachula, Mexico, using seven DNA microsatellites as molecular genetic markers. The mass-reared strains were a bisexual laboratory strain (LAB) with approximately 130 generations under mass-rearing and a genetic sexing strain, Tapachula-7 (TA7), also under mass-rearing for 100 generations. Our results revealed an overall low level of genetic differentiation (approximately 15%) among the three strains, with the LAB and WIL populations being genetically most similar and TA7 most genetically differentiated. Although there were some differences in allele frequencies between strains, our results show that overall, the adaptation to mass-rearing conditions did not reduce genetic variability compared to the wild sample in terms of heterozygosity or allelic richness, nor did it appear to alter the level of inbreeding with respect to the wild populations. These results are contrary to the general idea that mass-rearing always results in a reduction in genetic diversity. Overall, our findings can contribute to a better understanding of the impact that adaptation to mass-rearing conditions may have on the genetic make-up of strains.

Mitochondrial DNA and local ecological knowledge reveal two lineages of leatherback turtle on the beaches of Oaxaca, Mexico.

Despite multiple conservation efforts of the Mexican government, the leatherback turtle is at serious risk of extinction. In this study, we investigated the possible presence of a genetic bottleneck that could prevent the recovery of this species and compared these findings with those of the olive ridley turtle, which is in true recovery. Our results confirmed that a demographic change occurred in the past and the presence of two different leatherback turtle lineages that diverged approximately 13.5 million years ago. Local ecological knowledge (LEK) also described the presence of these two lineages and warned that one is at higher risk of extinction than the other. Genetic analysis confirmed 124 mutations between the two lineages, and much lower genetic diversity in one lineage than the other. Our study highlights and substantiates the power of mixing LEK, environmental history, and genetics to better understand conservation challenges of highly threatened species such as the leatherback turtle. Moreover, we report a new lineage of the leatherback turtle which may represent a distinct species. Future studies should focus on morphological, ecological, biogeographical, evolutionary and conservation perspectives for the analysis of the new lineage.

Does body size influence mating success? A morphometric study of two Anastrepha (Diptera: Tephritidae) fruit fly species.

Male mating success depends on various traits and factors, and correctly identifying these traits can be key in the context of pest management. For tephritid pests, controlled through the sterile insect technique (SIT) traits, such as male size, can be manipulated through mass-rearing procedures. Thus, it is particularly important to understand whether male size can favor mating success. Here, we evaluated mating success of males of different sizes in Anastrepha ludens and Anastrepha obliqua, two species controlled through SIT. For both species, a morphometric analysis was performed of mated and unmated mass-reared and wild males in field cages. In both A. ludens and A. obliqua, wild females did not discriminate their mates based on male size and mated more frequently with wild males regardless of size. For mass-reared males, we found no evidence of an advantage of the large males compared to small males in mating success. However, we did find differences between the morphometric traits of mass-reared and wild males. In A. ludens, traits associated to mating success were Face Width (FW), Head Width (HW), Thorax Length (TL) and Wing Length (WL), and for A. obliqua were FW, HW, WL and WW (Wing Width). Overall, FW and TL were more consistent predictors of mating success. In conclusion, female choice seems to suggest multivariate selection, confirming that overall body size (expressed as pupal diameter, which is highly correlated with weight) is not a decisive factor in male mating success in these two species. However, morphological traits such as FW, HW, TL, WL and WW may be relevant in mating preference of wild female.

How Does the Application of Beauveria bassiana and Compost on Corn Crops Affect the Survival and Genetic Diversity of Phyllophaga obsoleta (Coleoptera: Melolonthinae)?

The knowledge of the diversity and genetic structure of pest insects under management contributes to the improvement of control strategies. An experiment was run to investigate whether the addition of the fungus Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Cordycipitaceae) (BB) and compost to soil affects the presence and genetic diversity of adults and larvae of Phyllophaga obsoleta Blanch (Coleoptera: Melolonthinae) larvae in maize crops. We collected adults in and used mating pairs under four treatments (BB, compost, soil, blank). Genetic diversity and structure were determined through five allo/iso-enzymatic loci. Beauveria bassiana affected the presence and mortality of P. obsoleta in the laboratory but not under field conditions. The genetic diversity of P. obsoleta ranged from moderate to high (Ho = 0.26-0.31), with a low genetic differentiation among localities or treatments (Phi < 0.05), indicating high levels of gene flow. Our results showed a weak effect of B. bassiana on P. obsoleta in the field. Still, our laboratory observations suggest that the fungus may be a suitable alternative for biological control.

Genetic and Ecological Relationships of Anastrepha ludens (Diptera: Tephritidae) Populations in Southern Mexico.

Knowledge of the influence of evolutionary factors that promote either the differentiation or cohesion of pest insect populations is critical for the improvement of control strategies. Here, we explore the extent to which genetic differentiation occurs between populations of the Mexican fruit fly, Anastrepha ludens, in association with four plant hosts (Citrus sinensis, C. paradisi, Mangifera indica and Casimiroa edulis) in the Soconusco region of Chiapas (Mexico). Using variants from six enzymatic loci, we obtained measures of genetic diversity for three sample arrangements: (1) by sex per locality, (2) by locality and (3) by host. The extent of genetic differentiation in populations was assessed using the Analyses of Molecular Variance (AMOVA) method for each array of samples, and moderate to high levels of genetic variation were observed between the sexes, as well as among localities and host plants. A Bayesian approach was then used to assess any population structure underlying the genetic data we obtained, but this analysis showed no significant structuring due to locality or host plant. We also considered whether the observed genotypic frequencies in male and females matched those expected under a hypothesis of random mating. Here we found significant deviations from expected genotypic frequencies, suggesting that sexual selection is acting on these populations. Overall, our results indicate that sexual selection, along with the presence of some heterogeneity in environments provided by both geographical factors and availability of host plants, has influenced the evolution of pest populations in this region of Mexico. Implications for area-wide pest management strategies are discussed.

Mechanisms of diversity maintenance in dung beetle assemblages in a heterogeneous tropical landscape.

BACKGROUND: Anthropized landscapes play a crucial role in biodiversity conservation, as they encompass about 90% of the remaining tropical forest. Effective conservation strategies require a deep understanding of how anthropic disturbances determine diversity patterns across these landscapes. Here, we evaluated how attributes and assembly mechanisms of dung beetle communities vary across the Selva El Ocote Biosphere Reserve (REBISO) landscape. METHODS: Community attributes (species diversity, abundance, and biomass) were assessed at the landscape scale, using spatial windows and vegetation classes. Windows were categorized as intact, variegated, or fragmented based on their percent cover of tropical forest. The vegetation classes analyzed were tropical forest, second-growth forest, and pastures. RESULTS: We collected 15,457 individuals and 55 species. Variegated windows, tropical forests, and second-growth forests showed the highest diversity values, while the lowest values were found in intact windows and pastures. Landscape fragmentation was positively and strongly related to dung beetle diversity and negatively related to their abundance; biomass was positively associated with forest cover. Beta diversity was the primary driver of the high dung beetle diversity in the landscape analyzed. DISCUSSION: The landscape heterogeneity and its biodiversity-friendly matrix facilitate the complementarity of dung beetle assemblages in the Selva El Ocote Biosphere Reserve. Random processes govern beta diversity patterns in intact and variegated windows. Therefore, vegetation cover in the region is sufficient to maintain a continuous flow of dung beetles between forested landscape segments. However, intense anthropic disturbances acted as deterministic environmental filters in fragmented windows and pastures sites, leading to biotic homogenization processes. Our results suggest that increasing habitat variegation in highly fragmented sites is an effective strategy to prevent or buffer homogenization processes in the REBISO landscape.

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.

Distribución potencial y conectividad del paisaje: criterios para reevaluar el grado de amenaza de Campylorhynchus yucatanicus (Aves: Troglodytidae) / Potential distribution and landscape connectivity: criteria for reevaluating the threat degree of Campylorhynchus yucatanicus (Aves: Troglodytidae)

Resumen La distribución geográfica y la calidad del hábitat son criterios fundamentales para evaluar el grado de riesgo de extinción y amenaza de las especies. Campylorhynchus yucatanicus es un ave endémica de la costa norte de la Península de Yucatán, México con distribución restringida a una estrecha franja de hábitat, entre los estados de Campeche y Yucatán. En la actualidad, la costa yucateca ha perdido más de la mitad de la vegetación de duna costera y otros hábitats se han modificado principalmente por falta de un plan de urbanización y mal aprovechamiento de los recursos naturales. Estos factores amenazan a C. yucatanicus, que está catalogada como una especie casi amenazada por la UICN y como una especie en peligro de extinción por la legislación mexicana, NOM-059-2010. En este estudio se modeló su distribución potencial utilizando 64 registros de presencia de variadas fuentes (1960 y 2009), un conjunto de variables climáticas y una capa de Índice de vegetación de diferencia normalizada (NDVI). Para evaluar el grado de conectividad del paisaje se empleó un mapa de tipos de vegetación y uso de suelo, distancia a los poblados y a las carreteras asfaltadas. El modelo de distribución potencial arrojó una distribución potencial de 2 711 km2 aproximadamente, 2 % del área total de la Península de Yucatán. De esta área, solo el 27 % está protegido con categoría de Reserva de la Biósfera y sólo el 10 % corresponde a áreas núcleo de conservación, con restricciones de uso de suelo y protección relativamente efectiva. Las poblaciones de la región de Ría Lagartos y del oeste de Celestún, parecen ser las más aisladas, teniendo en cuenta el modelo de conectividad del paisaje. La permeabilidad del paisaje entre fragmentos de vegetación de duna cerca de la costa es baja, principalmente por la distribución de las zonas urbanas. Los resultados pueden ser usados para establecer estrategias de manejo y sugieren que la especie se encuentra en un estado de conservación más delicado del que hasta ahora se había descrito por la UICN. Consideramos que C. yucatanicus debe tener categoría en peligro de la UICN según su distribución y la conectividad de paisaje actual.

Abstract Geographic distribution and habitat quality are key criteria for assessing the degree of risk of species extinction threat. Campylorhynchusyucatanicus(Yucatán Wren, Troglodytidae) is an endemic bird of the Northern coast of the Yucatán Peninsula, Mexico, with a distribution restricted to a narrow strip of habitat, between Campeche and Yucatán states. Currently, the Yucatán coast has lost more than half of the coastal dune vegetation, and other habitats have been modified mainly because there is no urban development plan and the natural resources management is poor. These factors threaten C. yucatanicus, which is listed as a near threatened species by IUCN and as an endangered species by Mexican law NOM059-2010. In this paper, C. yucatanicus´s potential distribution was modeled using 64 presence records from several sources (1960 y 2009), a set of climate variables, and a vegetation index layer of normalized difference (NDVI). To assess the degree of landscape connectivity we used a map of vegetation types and land use, distance to villages and paved roads. The potential distribution model showed an area of approximately 2 711 km2, which is 2 % of the total area of the Yucatán Peninsula distribution. In this area, only 27 % is protected by Biosphere Reserve category and only 10 % belong to core conservation areas, with land use restrictions and relatively effective protection. The populations from Ría Lagartos and Western Celestún regions appear to be the most isolated following the model of landscape connectivity. Landscape permeability among fragments of dune vegetation near the coast is low, mainly due to the distribution of urban areas. These results can be used to establish management strategies, and show that the species is in more delicate conditions than what it has been described by IUCN. We consider that C. yucatanicus should be given endangered category by IUCN, because of their distribution and the context of the current landscape connectivity.

Fitness of Mass-Reared Males of Anastrepha obliqua (Diptera: Tephritidae) Resulting From Mating Competition Tests in Field Cages.

The sterile insect technique uses males that have been mass-reared in a controlled environment. The insects, once released in the field, must compete to mate. However, the mass-rearing condition supposes a loss of fitness that will be noticeable by wild females. To compare the fitness of wild males and mass-reared males, three competition settings were established. In setting 1, wild males, mass-reared males and wild females were released in field cages. In setting 2, wild females and wild males were released without competition, and in setting 3, mass-reared males and mass-reared females were also released without competition. Male fitness was based on their mating success, fecundity, weight and longevity. The fitness of the females was measured based on weight and several demographic parameters. The highest percentage of mating was between wild males and wild females between 0800 and 0900 h in the competition condition, while the mass-reared males started one hour later. The successful wild males weighed more and showed longer mating times, greater longevity and a higher number of matings than the mass-reared males. Although the mass-reared males showed the lowest percentage of matings, their fecundity when mating with wild females indicated a high fitness. Since the survival and fecundity of wild females that mated with mass-reared males decreased to become similar to those of mass-reared females that mated with mass-reared males, females seem to be influenced by the type of male (wild or mass-reared).

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.

Testing local host adaptation and phenotypic plasticity in a herbivore when alternative related host plants occur sympatrically.

Host race formation in phytophagous insects can be an early stage of adaptive speciation. However, the evolution of phenotypic plasticity in host use is another possible outcome. Using a reciprocal transplant experiment we tested the hypothesis of local adaptation in the aphid Brevicoryne brassicae. Aphid genotypes derived from two sympatric host plants, Brassica oleracea and B. campestris, were assessed in order to measure the extent of phenotypic plasticity in morphological and life history traits in relation to the host plants. We obtained an index of phenotypic plasticity for each genotype. Morphological variation of aphids was summarized by principal components analysis. Significant effects of recipient host on morphological variation and life history traits (establishment, age at first reproduction, number of nymphs, and intrinsic growth rate) were detected. We did not detected genotype × host plant interaction; in general the genotypes developed better on B. campestris, independent of the host plant species from which they were collected. Therefore, there was no evidence to suggest local adaptation. Regarding plasticity, significant differences among genotypes in the index of plasticity were detected. Furthermore, significant selection on PC1 (general aphid body size) on B. campestris, and on PC1 and PC2 (body length relative to body size) on B. oleracea was detected. The elevation of the reaction norm of PC1 and the slope of the reaction norm for PC2 (i.e., plasticity) were under directional selection. Thus, host plant species constitute distinct selective environments for B. brassicae. Aphid genotypes expressed different phenotypes in response to the host plant with low or nil fitness costs. Phenotypic plasticity and gene flow limits natural selection for host specialization promoting the maintenance of genetic variation in host exploitation.

Lifting a veil on diversity: a Bayesian approach to fitting relative-abundance models.

Bayesian methods incorporate prior knowledge into a statistical analysis. This prior knowledge is usually restricted to assumptions regarding the form of probability distributions of the parameters of interest, leaving their values to be determined mainly through the data. Here we show how a Bayesian approach can be applied to the problem of drawing inference regarding species abundance distributions and comparing diversity indices between sites. The classic log series and the lognormal models of relative- abundance distribution are apparently quite different in form. The first is a sampling distribution while the other is a model of abundance of the underlying population. Bayesian methods help unite these two models in a common framework. Markov chain Monte Carlo simulation can be used to fit both distributions as small hierarchical models with shared common assumptions. Sampling error can be assumed to follow a Poisson distribution. Species not found in a sample, but suspected to be present in the region or community of interest, can be given zero abundance. This not only simplifies the process of model fitting, but also provides a convenient way of calculating confidence intervals for diversity indices. The method is especially useful when a comparison of species diversity between sites with different sample sizes is the key motivation behind the research. We illustrate the potential of the approach using data on fruit-feeding butterflies in southern Mexico. We conclude that, once all assumptions have been made transparent, a single data set may provide support for the belief that diversity is negatively affected by anthropogenic forest disturbance. Bayesian methods help to apply theory regarding the distribution of abundance in ecological communities to applied conservation.