Alison P.M. Thomas

Avoidance of a generalist entomopathogenic fungus by the ladybird, Coccinella septempunctata

Fungal entomopathogens are ubiquitous within the environment and susceptible insects are predicted to be under strong selection pressure to detect and avoid virulent isolates. Beauveria bassiana is an entomopathogenic fungus with a wide host range including coccinellids. Seven-spot ladybirds, Coccinella septempunctata, overwinter predominantly in leaf litter and B. bassiana is one of their major mortality factors during winter in temperate regions. Behavioural assays were conducted to assess the ability of adult C. septempunctata to avoid lethal densities of B. bassiana conidia in soil or on leaves. Further assays considered avoidance by C. septempunctata of mycosed (B. bassiana) C. septempunctata cadavers compared with uninfected C. septempunctata cadavers or in vitro B. bassiana. Treatments in any bioassays entirely avoided by C. septempunctata were regarded as censored data, to overcome the difficulties associated with zeros in log-ratio analyses. Male and female C. septempunctata avoided contact with leaf surfaces and soil inoculated with B. bassiana and mycosed cadavers. The ability of C. septempunctata to detect and avoid B. bassiana conidia is an adaptation that undoubtedly increases survival and ultimately fitness. We predict that such behavioural responses are widespread and driven by the high cost of fungal infection to a host.

A fungal pathogen in time and space: the population dynamics of Beauveria bassiana in a conifer forest.

The fungal entomopathogen Beauveria bassiana is ubiquitous in below-ground systems; however, there is a dearth of information on the above-ground diversity, temporal and spatial distribution of this fungus. Therefore, we assessed its occurrence in a conifer forest (Pseudotsuga monziesii and Pinus nigra var. maritima) using selective media to isolate B. bassiana from soil, branch and bark samples collected in October 2005, March and June 2006. Fungal density was the highest at all locations in October, declining in March and June, and absent from conifer branches in June. This above-ground decline most likely resulted from more extreme environmental conditions compared with those below ground. Molecular analyses (ISSR-PCR) indicated that B. bassiana is genetically diverse, comprising both distinct microhabitat-specific and seasonal isolates. The occurrence of dissimilar above- and below-ground isolates suggests that B. bassiana occupies various overlapping niches in these systems.

Predation of native coccinellids by the invasive alien Harmonia axyridis (Coleoptera: Coccinellidae): detection in Britain by PCR-based gut analysis

Abstract.  1. The harlequin ladybird, Harmonia axyridis, is an invasive alien species that arrived in Britain in 2003 and has spread across most of the country.

The Genetic Basis of Melanism in the Gray Squirrel (Sciurus carolinensis)

The black squirrel is a melanic variant of the gray squirrel (Sciurus carolinensis). We found 3 coat color variants in the gray squirrel: the wild-type gray, a jet-black, and a brown-black phenotype. These 3 morphs are due to varying distributions of eumelanin and phaeomelanin pigment in hairs. The melanocortin 1 receptor (MC1R) plays a central role in regulating eumelanin and phaeomelanin production. We sequenced the MC1R gene for all 3 coat color phenotypes and found a 24 base-pair deletion. The gray phenotype was homozygous for the wild-type allele E(+), the jet-black phenotype was homozygous for the MC1R-Delta24 allele E(B), and the brown-black phenotype was heterozygous for the E(+) and E(B) alleles. We conclude that melanism in gray squirrels is associated with the MC1R-Delta24 E(B) allele at amino acid positions 87-94 and that this allele is incompletely dominant to the wild-type allele. We predict that the MC1R-Delta24 E(B) allele encodes a constitutively active or hyperactive receptor.

Intraguild predation by Harmonia axyridis (Coleoptera: Coccinellidae) on native insects in Europe: molecular detection from field samples

Intraguild predation (IGP) is a potential mechanism of negative effects on native species populations by invasive non‐native species such as Harmonia axyridis. Molecular techniques (polymerase chain reaction) were used to probe for the presence of various insect (coccinellid, syrphid and chrysopid) prey DNA in the guts of 177 H. axyridis larvae field‐collected in England, France, Germany, Slovakia and the Czech Republic in 2010. Three of the four target prey species were detected in the guts of H. axyridis at the following rates: Adalia decempunctata 9.6%, Adalia bipunctata 2.8% and Episyrphus balteatus 2.8%. IGP on Chrysoperla carnea was not detected. IGP detection of at least one target species was made in England, France, Slovakia and the Czech Republic, but not in Germany. These results strengthen the evidence that H. axyridis is a very generalist predator. Results are discussed in the context of their ecological significance.

Detecting arthropod intraguild predation in the field

The process of biological control carries a distinct risk that an alien biological control agent (BCA) will become established as an invasive alien species with an associated threat to the local ecosystem biodiversity. It is imperative that a wide-ranging environmental risk assessment (ERA) is performed before the release of any BCA. This should include considering various potential but difficult to observe ecological interactions between the BCA and members of the native community, including disruption of intraguild relationships. Detection of intraguild predation (IGP) events involving predatory arthropods in the field can be done by analyzing their gut contents. Polymerase chain reaction (PCR) is a sensitive and specific tool to identify target prey DNA within a predator’s gut. This paper reviews the efficiency of a DNA based approach for detecting IGP in the field, compared with detection by the use of monoclonal antibodies or gas chromatography. Prey specificity, detection times after prey consumption, capacity for quantification, multiple prey targeting and the time and costs involved in developing and using the different methods are considered.

Presence of haemocyte-like cells in coccinellid reflex blood

Abstract.  Contrary to current assumptions, the reflex blood of two‐spot ladybirds, Adalia bipunctata, and seven‐spot ladybirds, Coccinella septempunctata, contains haemocyte‐like cells. Furthermore, DNA can be extracted and amplified from coccinellid reflex blood, confirming the presence of haemocyte‐like cells and demonstrating a nondestructive method of DNA extraction.

Melanocortin 1 Receptor (MC1R) Gene Sequence Variation and Melanism in the Gray (Sciurus carolinensis), Fox (Sciurus niger), and Red (Sciurus vulgaris) Squirrel

Sequence variations in the melanocortin 1 receptor (MC1R) gene are associated with melanism in many different species of mammals, birds, and reptiles. The gray squirrel (Sciurus carolinensis), found in the British Isles, was introduced from North America in the late 19th century. Melanism in the British gray squirrel is associated with a 24-bp deletion in the MC1R. To investigate the origin of this mutation, we sequenced the MC1R of 95 individuals including 44 melanic gray squirrels from both the British Isles and North America. Melanic gray squirrels of both populations had the same 24-bp deletion associated with melanism. Given the significant deletion associated with melanism in the gray squirrel, we sequenced the MC1R of both wild-type and melanic fox squirrels (Sciurus niger) (9 individuals) and red squirrels (Sciurus vulgaris) (39 individuals). Unlike the gray squirrel, no association between sequence variation in the MC1R and melanism was found in these 2 species. We conclude that the melanic gray squirrel found in the British Isles originated from one or more introductions of melanic gray squirrels from North America. We also conclude that variations in the MC1R are not associated with melanism in the fox and red squirrels.