A PCR-based tool for the cultivation-independent monitoring of Pandora neoaphidis.

Pandora neoaphidis is one of the most important fungal pathogens of aphids and has a great potential for use in biocontrol. Little is known on how this fungus persists in an area and in particular on its overwintering strategies. It is hypothesized that natural areas play an important role for survival and that soil may serve as a source of inoculum for new aphid populations in spring. To test these hypotheses, a cultivation-independent PCR-based diagnostic tool was developed, that allows the detection of P. neoaphidis in the environment. Two P. neoaphidis specific PCR primer pairs were designed, targeting sequences in the ribosomal RNA gene cluster. Specificity of both primer pairs was demonstrated with P. neoaphidis and non-target close entomophthoralean relatives. Moreover, single amplicons of expected sizes were obtained with both primer pairs from various environmental sample types, including aphid cadavers, plant material, and soil. The PCR-based diagnostic tool was applied to investigate the persistence of P. neoaphidis in soil samples obtained in 2004/2005 from a nettle field harboring infected aphids in fall 2004. P. neoaphidis was detected in every sample collected in November 2004 and March 2005, suggesting an overwintering stage of P. neoaphidis in top soil layers. The developed cultivation-independent PCR-based tool will be valuable for further investigation of the ecology of P. neoaphidis and for the development and future implementation of management strategies against aphids involving conservation biocontrol.

Invasive zygomycosis: update on pathogenesis, clinical manifestations, and management.

Zygomycosis is an increasingly common infection in immunocompromised patients. Advances in the understanding of Zygomycetes pathobiology and the introduction of new drugs with improved activity and tolerability for treatment of zygomycosis have improved the prospects of effectively controlling this devastating infection. Further reductions in mortality will require improved diagnostic and novel therapeutic approaches for this group of aggressive opportunistic molds.

Detection of villous conidia of Conidiobolus coronatus in a blood sample by scanning electron microscopy investigation.

Conidiobolus coronatus is a major insect pathogen belonging to the fungal order Entomophthorales, causing a rare subcutaneous infection of the nasofacial region, resulting in swelling of predominantly the nose, mouth, and perinasal tissue. Later in the course of the infection firm, painless, subcutaneous nodules develop that are attached to the underlying tissues but not to the skin. No morphological studies are available in the literature on the morphology of C. coronatus in vivo and all morphological studies have been conducted on in vitro cultures. Here the authors report on the ultrastructural pathology as seen with a scanning electron microscope (SEM) of villous conidia of C. coronatus, detected in a 37-year-old woman who presented to the casualty department at Pretoria Academic Hospital, South Africa with left-sided facial pain and headache. The diagnosis of C. coronatus was confirmed by LightCycler real-time flourescence PCR technique. Research shows that typically diagnosis of the pathogen is established only on histological examination, and in over 85% of cases cultures for the causative organism is negative. This pathogen has not previously been found in a blood sample and the authors present for the first time the morphology of C. coronatus in blood using the SEM.