Beyond the Limits of Light: An Application of Super-Resolution Confocal Microscopy (sCLSM) to Investigate Eocene Amber Microfossils.

Amber is known as one of the best sources of fossil organisms preserved with exceptional fidelity. Historically, different methods of imaging have been applied to amber, including optical microscopy and microtomography. These methods are sufficient to resolve millimeter-scaled fossils. However, microfossils, such as microarthropods, require another resolution. Here, we describe a non-destructive method of super resolution confocal microscopy (sCLSM) to study amber-preserved microfossils, using a novel astigmatid mite species (genus Histiogaster, Acaridae) from Eocene Rovno amber as a model. We show that the resolution obtained with sCLSM is comparable to that of scanning electron microscopy (SEM) routinely used to study modern mites. We compare sCLSM imaging to other methods that are used to study amber inclusions and emphasize its advantages in examination of unique fossil specimens. Furthermore, we show that the deterioration of amber, which manifests in its darkening, positively correlates with its increased fluorescence. Our results demonstrate a great potential of the sCLSM method for imaging of the tiniest organisms preserved in amber.

Flexibility in the ambrosia symbiosis of Xyleborus bispinatus.

Introduction: Ambrosia beetles maintain strict associations with specific lineages of fungi. However, anthropogenic introductions of ambrosia beetles into new ecosystems can result in the lateral transfer of their symbionts to other ambrosia beetles. The ability of a Florida endemic ambrosia beetle, Xyleborus bispinatus, to feed and establish persistent associations with two of its known symbionts (Raffaelea subfusca and Raffaelea arxii) and two other fungi (Harringtonia lauricola and Fusarium sp. nov.), which are primary symbionts of invasive ambrosia beetles, was investigated. Methods: The stability of these mutualisms and their effect on the beetle's fitness were monitored over five consecutive generations. Surface-disinfested pupae with non-developed mycangia were reared separately on one of the four fungal symbionts. Non-treated beetles (i.e., lab colony) with previously colonized mycangia were used as a control group. Results: Xyleborus bispinatus could exchange its fungal symbionts, survive, and reproduce on different fungal diets, including known fungal associates and phylogenetically distant fungi, which are plant pathogens and primary symbionts of other invasive ambrosia beetles. These changes in fungal diets resulted in persistent mutualisms, and some symbionts even increased the beetle's reproduction. Females that developed on Fusarium sp. nov. had a significantly greater number of female offspring than non-treated beetles. Females that fed solely on Harringtonia or Raffaelea symbionts produced fewer female offspring. Discussion: Even though some ambrosia beetles like X. bispinatus can partner with different ambrosia fungi, their symbiosis under natural conditions is modulated by their mycangium and possibly other environmental factors. However, exposure to symbionts of invasive beetles can result in stable partnerships with these fungi and affect the population dynamics of ambrosia beetles and their symbionts.

Macrochelid mites (Mesostigmata: Macrochelidae) from São Paulo state, Brazil, with description of a new species of Macrocheles.

Macrochelid mites have been reported in several papers about mites of different habitats in Brazil, but this is the first work specifically conducted to determine the macrochelids from that country. The main goal was to determine macrochelid species in microhabitats where the stable fly is usually found in commercial cattle farms. In total, 1359 specimens were collected from cow manure and litter in different regions of São Paulo state. These represented ten species of three genera. One of these, Macrocheles embersoni n. sp. is here described as new to science. Other macrochelid species are expected to be found in the study area, when other types of microhabitats are explored.