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Pesticides use raises concerns regarding environmental sustainability, as pesticides are closely linked to the decline of biodiversity and adverse human health outcomes. This study proposed a holistic approach for assessing the potential risks posed by pesticides for human health and the environment in the eastern region of Michoacan, where extensive agricultural lands, especially corn and avocado fields, surround the Monarch Butterfly Biosphere Reserve. We used a combination of qualitative (semi-structured interviews) and quantitative (chemical analysis) data. Fifty-five interviews with smallholder farmers allowed us to identify pesticide types, quantities, frequencies, and application methods. A robust and precise analytical method based on solid-phase extraction and LC-MS/MS was developed and validated to quantify 21 different pesticides in 16 water samples (rivers, wells, runoff areas). We assessed environmental and human health risks based on the pesticides detected in the water samples and reported in the interviews. The interviews revealed the use of 28 active ingredients, including glyphosate (29 % of respondents), imidacloprid (27 %), and benomyl (24 %). The pesticide analysis showed the presence of 13 different pesticides and degradation products in the water samples. The highest concentrations were found for imidacloprid (1195 ngL-1) and carbendazim (a degradation product of benomyl; 932 ngL-1), along with the metabolite of pyrethroid insecticides, 3-PBA (494 ngL-1). The risk assessment indicates that among the most used pesticides, the fungicide benomyl and carbendazim pose the highest risk to human health and aquatic ecosystems, respectively. This study unveils novel insights on agricultural practices for the avocado, a globally consumed crop that is undergoing rapid production expansion. It calls for the harmonisation of crop protection with environmental responsibility, safeguarding the health of the people involved and the surrounding ecosystems.

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Monarch butterflies (Danaus plexippus) are well studied for their annual long-distance migration from as far north as Canada to their overwintering grounds in Central Mexico. At the end of the cold season, monarchs start to repopulate North America through short-distance migration over the course of multiple generations. Interestingly, some populations in various tropical and subtropical islands do not migrate and exhibit heritable differences in wing shape and size, most likely an adaptation to island life. Less is known about forewing differences between long- and short-distance migrants in relation to island populations. Given their different migratory behaviors, we hypothesized that these differences would be reflected in wing morphology. To test this, we analyzed forewing shape and size of three different groups: nonmigratory, lesser migratory (migrate short-distances), and migratory (migrate long-distances) individuals. Significant differences in shape appear in all groups using geometric morphometrics. As variation found between migratory and lesser migrants has been shown to be caused by phenotypic plasticity, and lesser migrants develop intermediate forewing shapes between migratory and nonmigratory individuals, we suggest that genetic assimilation might be an important mechanism to explain the heritable variation found between migratory and nonmigratory populations. Additionally, our research confirms previous studies which show that forewing size is significantly smaller in nonmigratory populations when compared to both migratory phenotypes. Finally, we found sexual dimorphism in forewing shape in all three groups, but for size in nonmigratory populations only. This might have been caused by reduced constraints on forewing size in nonmigratory populations.

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Monarch butterflies are known for their spectacular annual migration in eastern North America, with millions of monarchs flying up to 4,500 km to overwintering sites in central Mexico. Monarchs also live west of the Rocky Mountains, where they travel shorter distances to overwinter along the Pacific Coast. It is often assumed that eastern and western monarchs form distinct evolutionary units, but genomic studies to support this notion are lacking. We used a tethered flight mill to show that migratory eastern monarchs have greater flight performance than western monarchs, consistent with their greater migratory distances. However, analysing more than 20 million SNPs in 43 monarch genomes, we found no evidence for genomic differentiation between eastern and western monarchs. Genomic analysis also showed identical and low levels of genetic diversity, and demographic analyses indicated similar effective population sizes and ongoing gene flow between eastern and western monarchs. Gene expression analysis of a subset of candidate genes during active flight revealed differential gene expression related to nonmuscular motor activity. Our results demonstrate that eastern and western monarchs maintain migratory differences despite ongoing gene flow, and suggest that migratory differences between eastern and western monarchs are not driven by select major-effects alleles. Instead, variation in migratory distance and destination may be driven by environmentally induced differential gene expression or by many alleles of small effect.

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Every year, Danaus plexippus (Linnaeus, 1758) travels to hibernate in oyamel fir forests located between the limits of the states of Michoacán and Mexico in Mexico. Climate change and anthropogenic actions are diminishing oyamel fir forests in Mexico, putting pressure on the habitats of monarch butterflies. In the last decade, new colonies outside their usual range have been predicted through modeling and reported by the National Commission on Protected Areas of Mexico. The objectives of the study were to recover information on the historical and new hibernation sites, reported or modeled, from different literature sources. We also aimed to perform a bioclimatic and forest biometric characterization of new monarch butterfly colonies located in Sierra Nevada in Mexico to provide information to aid in conservation strategies for the monarch butterfly population. We conducted field trips to georeference the colonies at sites located in the Atlautla municipality in Mexico State. Climatic, topographic, and forest biometric variables were used to characterize the sites physically. It was found that the butterfly's roosts occurred at a higher elevation than those recorded by other sources. The locations where the monarch's colonies were established, in the east of Mexico State, provide information relevant to defining and developing policies for their conservation.

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Abies religiosa forests in central Mexico are the only overwinter refuge of the monarch butterfly and provide important ecosystem services. These forests have lost 55% of their original area and as a consequence, diversity and biotic interactions in these ecosystems are in risk. The aim of this study was to compare the soil fungal diversity and community structure in the Abies religiosa forests and surrounding Pinus montezumae, Pinus hartwegii, and coniferous mixed forest plant communities to provide data on ecology of mycorrhizal interactions for the assisted migration of A. religiosa. We sampled soil from five coniferous forests, extracted total soil DNA, and sequenced the ITS2 region by Illumina MiSeq. The soil fungi community was integrated by 1746 taxa with a species turnover ranging from 0.280 to 0.461 between sampling sites. In the whole community, the more abundant and frequent species were Russula sp. (aff. olivobrunnea), Mortierella sp.1, and Piloderma sp. (aff. olivacearum). The ectomycorrhizal fungi were the more frequent and abundant functional group. A total of 298 species (84 ectomycorrhizal) was shared in the five conifer forests; these widely distributed species were dominated by Russulaceae and Clavulinaceae. The fungal community composition was significantly influenced by altitude and the lowest species turnover happened between the two A. religiosa forests even though they have different soil types. As Pinus montezumae forests have a higher altitudinal distribution adjacent to A. religiosa and share the largest number of ectomycorrhizal fungi with it, we suggest these forests as a potential habitat for new A. religiosa populations.

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Given the rapid population decline and recent petition for listing of the monarch butterfly (Danaus plexippus L.) under the Endangered Species Act, an accurate estimate of the Eastern, migratory population size is needed. Because of difficulty in counting individual monarchs, the number of hectares occupied by monarchs in the overwintering area is commonly used as a proxy for population size, which is then multiplied by the density of individuals per hectare to estimate population size. There is, however, considerable variation in published estimates of overwintering density, ranging from 6.9-60.9 million ha-1. We develop a probability distribution for overwinter density of monarch butterflies from six published density estimates. The mean density among the mixture of the six published estimates was ∼27.9 million butterflies ha-1 (95% CI [2.4-80.7] million ha-1); the mixture distribution is approximately log-normal, and as such is better represented by the median (21.1 million butterflies ha-1). Based upon assumptions regarding the number of milkweed needed to support monarchs, the amount of milkweed (Asclepias spp.) lost (0.86 billion stems) in the northern US plus the amount of milkweed remaining (1.34 billion stems), we estimate >1.8 billion stems is needed to return monarchs to an average population size of 6 ha. Considerable uncertainty exists in this required amount of milkweed because of the considerable uncertainty occurring in overwinter density estimates. Nevertheless, the estimate is on the same order as other published estimates. The studies included in our synthesis differ substantially by year, location, method, and measures of precision. A better understanding of the factors influencing overwintering density across space and time would be valuable for increasing the precision of conservation recommendations.

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Community-based small-scale reforestation practices have been proposed as an alternative to low-efficiency massive reforestations conducted by external agents. These latter conventional reforestations are often carried out in soils that have been seriously degraded and this has indirectly contributed to the introduction of non-native species and/or acceptance of very low seedling survival rates. Bokashi is a fermented soil organic amendment that can be made from almost any available agricultural byproduct, and its beneficial effects in agriculture have been reported in various contexts. Here, we report the results of a community-based small-scale experimental reforestation where the provenance of pine seedlings (local and commercial) and the use of Bokashi as a soil amendment were evaluated. Bokashi was prepared locally by members of a small rural community in central Mexico. Almost two years after the establishment of the trial, survival rates for the unamended and amended local trees were 97-100% while survival of the commercial trees from unamended and amended treatments were 87-93%. Consistently through time, local and commercial seedlings planted in Bokashi-amended soils were significantly taller (x̅ = 152 cm) than those planted in unamended soils (̅x = 86 cm). An unplanned infection by Cronartium quercuum in the first year of the experiment was considered as a covariable. Infected seedlings showed malformations but this did not affect survival and growth rates. Bokashi amendment seems as an inexpensive, locally viable technology to increase seedling survival and growth and to help recover deforested areas where soils have been degraded. This allows local stakeholders to see more rapid results while helping them to maintain their interest in conservation activities.

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Forest conservation plays a significant role in environmental sustainability. In Mexico only 8.48 million ha of forest are used for conservation of biodiversity. Payment for Environmental Services in the Monarch Butterfly Biosphere Reserve, one of the most important national protected areas, contributes to the conservation of these forests. In the Reserve, production of rainbow trout has been important for the rural communities who need to conserve the forest cover in order to maintain the hibernation cycle of the butterfly. Aquaculture is a highly productive activity for these protected areas, since it harnesses the existing water resources. In this study, changes from 1999 to 2012 in vegetation and land-use cover in the El Lindero basin within the Reserve were evaluated in order to determine the conservation status and to consider the feasibility of aquaculture as a means of sustainable development at community level. Evaluation involved stereoscopic interpretation of digital aerial photographs from 1999 to 2012 at 1:10,000 scale, comparative analysis by orthocorrected mosaics and restitution on the mosaics. Between 1999 and 2012, forested land recovered by 28.57 ha (2.70%) at the expense of non-forested areas, although forest degradation was 3.59%. Forest density increased by 16.87%. In the 46 ha outside the Reserve, deforestation spread by 0.26%, and land use change was 0.11%. The trend towards change in forest cover is closely related to conservation programmes, particularly payment for not extracting timber, reforestation campaigns and surveillance, whose effects have been exploited for the development of rural aquaculture; this is a new way to improve the socio-economic status of the population, to avoid logging and to achieve environmental sustainability in the Reserve.

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With international concern about the rates of deforestation worldwide, particular attention has been paid to Latin America. Forest conservation programmes in Mexico include Payment for Environmental Services (PES), a scheme that has been successfully introduced in the Monarch Butterfly Biosphere Reserve. To seek further evidence of the role of PES in lessening land degradation processes in a temperate region, the conservation state of the Cerro Prieto ejido within the Reserve was assessed by an analysis of changes in vegetation cover and land-use between 1971 and 2013. There were no changes in the total forest surface area, but the relative proportions of the different classes of cover density had changed. In 1971, closed and semi-closed forest occupied 247.81 ha and 5.38 ha, 82.33% and 1.79% of the total area of the ejido, respectively. By 2013, closed forest had decreased to 230.38 ha (76.54% of the ejido), and semi-closed cover was 17.23 ha (5.72% of the ejido), suggesting that some semi-closed forest had achieved closed status. The final balance between forest losses and recovery was: 29.63 ha were lost, whereas 13.72 ha were recovered. Losses were mainly linked to a sanitation harvest programme to control the bark beetle Scolytus mundus. Ecotourism associated with forest conservation in the Cerro Prieto ejido has been considered by inhabitants as a focal alternative for economic development. Consequently, it is essential to develop a well-planned and solidly structured approach based on social cohesion to foster a community-led sustainable development at local level.

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