A survey of possible associations between preharvest environment conditions and postharvest rejections of cut freesia flowers

Botrytis cinerea is the major pathogen infecting cut freesia flowers. Flecking symptoms on petals caused by this fungus result in postharvest rejections and substantial economic loss to both growers and sellers. In a limited survey for industry, numbers of freesia stems sent from a specialist grower in The Netherlands and rejected at a cut flower wholesaler in the United Kingdom were documented. Relationships between preharvest environment conditions in Holland that may predispose flowers to infection and postharvest freesia rejection levels in the United Kingdom due to B. cinerea flecking symptom expression are reported. Freesia rejections peaked during spring and, to a lesser degree, autumn periods. However, no clear correlations between preharvest growing environment conditions (e.g. 3-day means for temperature preceding harvest) and postharvest rejection frequency (%) could be discerned. Thus, sporadic freesia rejections in the United Kingdom were probably attributable either to other unresolved variables during the pre- (e.g. infection pressure) and/or postharvest (e.g. condensation events) phases or to interactions among predisposing variables.

A randomised controlled study of the efficacy of hypromellose and Lacri-Lube combination versus polyethylene/Cling wrap to prevent corneal epithelial breakdown in the semiconscious intensive care patient

Objective. To compare the efficacy of two forms of eye care (hypromellose and Lacri-Lube combination vs polyethylene/Cling wrap covers) for intensive care patients. Design. Randomised-controlled trial. Setting. University affiliated, tertiary referral hospital. Patients and participants. One hundred ten patients with a reduced or absent blink reflex were followed through until they regained consciousness, were discharged from the facility during study enrolment, died or developed a positive corneal ulcer or eye infection. Interventions. All patients received standard eye cleansing every 2 h. In addition to this, group one (n=60) received a treatment combining hypromellose drops and Lacri-Lube (HL) to each eye every 2 h. Group two (n=50) had polyethylene covers only placed over the eye to create a moisture chamber. Measurements and results. Corneal ulceration was determined using corneal fluorescein stains and mobile slit lamp evaluation, performed daily. No patients had corneal ulceration in the polyethylene cover group, but 4 patients had corneal ulceration in the HL group. Conclusions. Polyethylene covers are as effective as HL in reducing the incidence of corneal damage in intensive care patients.

A second Candida albicans resistance gene (Carg2) regulates tissue damage, but not fungal clearance, in sub-lethal murine systemic infection

The severity of systemic infection with the yeast Candida albicans has been shown to be under complex genetic control. C57/L mice carry an allele that is associated with an increase in tissue destruction when compared with C57BI/6 mice; however, the gene affects only the severity of tissue lesions, and does not influence the magnitude of the fungal burden in either kidney or brain. Studies in [C57/L x C57BI/6]F1 hybrid mice, and [C57/L x C57BI/6]F1 x C57/L backcross mice, demonstrated that the gene behaves as a simple Mendelian co-dominant. (C) 1998 Academic Press.

Type 1 and Type 2 cytokines: from basic science to fungal infections

At the present time, it is clear that Th1 responses afford protection against the fungi; however, the development, maintenance and function of the protective immune responses are complex mechanisms and are influenced by multiple factors. The route of infection has been shown to affect initial cytokine production and, consequently, the induction of protective Th1 responses. The ability of different isolates of the same fungal agent to induce and sustain a protective response has also been emphasized. Protective immune responses have been shown to vary in genetically different mouse strains after infection. In addition, these protective responses, such as cellular influx and cytokine production, also vary within the same animal depending on the tissue infected. The functional dominance of certain cytokines over others in influencing development and maintenance of protective responses has been discussed. Certain cytokines may act differently in hosts lacking important components of their innate or immune repertoire. It is evident from these presentations that a more comprehensive understanding of the protective mechanisms against different fungal agents is emerging. However, there is still much to learn before cytokine modulatory therapy can be used effectively without risk in the human host.

Acute susceptibility of aged mice to infection with Candida albicans

The effect of aging on host resistance to systemic candidosis was assessed by monitoring the course of infection in 16-month-old CBA/CaH mice (aged non-immune) and in a comparable group that had been infected with a sublethal dose of Candida albicans at 6 weeks of age (aged immune). Aged non-immune mice showed rapid progression of the disease, with a marked increase in the number of mycelia in the brain and kidney, and early morbidity, Foci of myocardial necrosis were evident, but inflammatory cells were sparse. The histological picture in the aged immune mice was similar to that in the aged non-immune group, although fewer mycelial aggregates were seen. Both groups of aged mice showed a significantly lower fungal burden in the brain on day 1 of infection, but on day 4, colony counts increased significantly in the aged non-immune mice, Comparison of cytokine gene expression in the infected brains showed that the relative amount of interferon-gamma and tumour necrosis factor-alpha cDNA were similar in all three groups. Interleukin-6 was elevated in both infected non-immune and uninfected aged mice. Aged immune mice showed no morbidity after challenge, and both colonisation and tissue damage were reduced in comparison with the aged non-immune animals.

Infection of the immature stages of Diadegma semiclausum, an endolarval parasitoid of the diamondback moth, by Beauveria bassiana

Interactions between the immature stages of Diadegma semiclausum, an endolarval parasitoid of Plutella xylostella, and the fungal entomopathogen Beauveria bassiana were investigated in the laboratory. Detrimental effects of B. bassiana on D. semiclausum cocoon production and adult parasitoid emergence increased with increasing pathogen concentration and some parasitoid larvae became infected by B. bassiana within hosts. The negative impact of B. bassiana on D. semiclausum cocoon production decreased as temporal separation between parasitism and pathogen exposure increased. Adult parasitoid emergence was significantly compromised by the highest rates of B. bassiana tested even when exposure of host larvae to the pathogen was delayed until one day before predicted parasitoid cocoon formation. Parasitoid pupae were infected by the pathogen in all B. bassiana treatments which did not preclude their development. (C) 2004 Elsevier Inc. All rights reserved.

Initiation of fungal epizootics in diamondback moth populations within a large field cage: proof of concept for auto-dissemination

Adult diamondback moths (DBM), Plutella xylostella L. (Lepidoptera: Plutellidae), inoculated with the fungus Zoophthora radicans, were released within a large field cage containing DBM-infested potted broccoli plants. Larvae and pupae on exposed and caged control plants were examined on five occasions over the next 48 days for evidence of Z. radicans infection. Infected larvae were first detected on exposed plants 4 days after the initial release of adults, and after 48 days the infection level reached 79%. Aerially borne conidia were a factor in transmission of the fungus. Infection had no effect on possible losses of larval and adult cadavers due to scavengers in field crops. In a trial to measure the influence of infection on dispersal, twice as many non-infected as infected males were recaptured in pheromone traps, although the difference in cumulative catch only became significant 3 days after release of the males. In a separate experiment, when adult moths were inoculated with Beauveria bassiana conidia and released into the field cage, DBM larvae collected from 37 of 96 plants sampled 4 days later subsequently died from B. bassiana infection. The distribution of plants from which the infected larvae were collected was random, but the distribution of infected larvae was clustered within the cage. These findings suggest that the auto-dissemination of fungal pathogens may be a feasible strategy for DBM control, provided that epizootics can be established and maintained when DBM population densities are low.

Disseminated Scedosporium prolificans infection and survival of a child with acute lymphoblastic leukemia

Scedosporium prolificans is a saprophytic fungus responsible for an increasing number of infections among immumocompromised hosts. Historically, disseminated infection with this organism has resulted in death. We report on a pediatric patient who developed overwhelming S. prolificans sepsis after induction chemotherapy for acute lymphoblastic leukemia. She is well 18 months after the diagnosis of fungal sepsis and continues to receive chemotherapy for leukemia, which remains in remission.

Active and passive immunization against oral Candida albicans infection in a murine model

Background/aims: Clinical and laboratory studies are consistent with a major role for cell-mediated immunity in recovery from oral infection with Candida albicans, but the role of humoral immunity remains controversial. The purpose of this study was to establish the relative contributions of cellular and humoral immunity to protection against oral candidiasis in a murine model, and to determine whether host responses could be enhanced by different immunization strategies. Results: Active oral immunization was protective in BALB/c and CBA/CaH mice, reducing both fungal burden and duration of infection after secondary challenge, whereas systemic immunization failed to protect against subsequent oral challenge. Candida-specific IgM was the predominant antibody detected in serum following both primary and secondary oral challenge; however, Candida-specific salivary IgA was not detectable. Immunization by passive transfer of either lymphocytes or immune serum did not confer any significant protection against oral infection in either susceptible or resistant mouse strain. Conclusion: The data demonstrate a possible role for mucosa-associated immunity following active immunization by the oral route, most likely exerted by local T lymphocytes resident in the oral mucosa, but there was no evidence to support a role for humoral immunity in protection against oral candidiasis.

CgDN24: A gene involved in hyphal development in the fungal phytopathogen Colletotrichum gloeosporioides

A cDNA corresponding to a transcript induced in culture by N starvation, was identified in Colletotrichum gloeosporioides by a differential hybridisation strategy. The cDNA comprised 905 bp and predicted a 215 aa protein; the gene encoding the cDNA was termed CgDN24. No function for CgDN24 could be predicted by database homology searches using the cDNA sequence and no homologues were found in the sequenced fungal genomes. Transcripts of CgDN24 were detected in infected leaves of Stylosanthes guianensis at stages of infection that corresponded with symptom development. The CgDN24 gene was disrupted by homologous recombination and this led to reduced radial growth rates and the production of hyphae with a hyperbranching phenotype. Normal sporutation was observed, and following conidia inoculation of S. guianensis, normal disease development was obtained. These results demonstrate that CgDN24 is necessary for normal hyphal development in axenic culture but dispensable for phytopathogenicity. (c) 2005 Elsevier GmbH. Alt rights reserved.

Postharvest infection of Freesia hybrida flowers by Botrytis cinerea

'Specking' on harvested freesia (Freesia hybrida) flowers is a problem worldwide. The disease is caused by the fungal pathogen Botrytis cinerea. This disease symptom detracts from appearance and reduces marketability of the flowers. Unlike other important cut flower crops (e.g. gerbera), the mode of infection and epidemiology of postharvest freesia flower specking caused by B. cinerea has not been reported. Epidemiological studies were carried out under simulated conditions typical of those occurring during postharvest handling of freesia flowers. Infection of freesia flowers by B. cinerea occurred when a conidium germinated, formed a germ tube(s) and penetrated epidermal cells. Fungal hyphae then colonised adjacent cells, resulting in visible lesions. Different host reactions were observed on freesia 'Cote d'Azur' petals at 20 degrees C compared to 5 degrees C. The infection process was relatively rapid at 20 degrees C, with visible lesions produced within 7 h of incubation. However, lesion expansion ceased after 24 h of incubation. Infection was slower at 5 degrees C, with visible lesions produced after 48 h of incubation. However, lesion development at 5 degrees C was continuous, with lesions expanding over 4 days. Light microscopy observations revealed increased host defence reactions during infection. These reactions involved production of phenolic compounds, probably lignin and/or callose, around infection sites. Such substances may play a role in restricting petal colonisation and lesion expansion. Disease severity and lesion numbers on freesia flowers incubated at 12 degrees C were higher, but not significantly higher (P > 0.05), than on those incubated at 20 degrees C. Disease severity and progression were differentially mediated by temperature and relative humidity (R. H.). Infection of freesia flowers was severe at 100% R. H. for all three incubation temperatures of 5, 12 and 20 degrees C. In contrast, no lesions were produced at 80 to 90% R. H. at either 5 or 20 degrees C.

Wolbachia pipientis: intracellular infection and pathogenesis in Drosophila

Wolbachia pipientis is a vertically transmitted, obligate intracellular symbiont of arthropods. The bacterium is best known for its ability to manipulate host reproductive biology where it can induce cytoplasmic incompatibility, parthenogenesis, feminization and male-killing. In addition to the various reproductive phenotypes it generates through interaction with host reproductive tissue it is also known to infect somatic tissues. However, relatively little is known about the consequences of infection of these tissues with the exception that in some hosts Wolbachia acts as a classical mutualist and in others a pathogen, dramatically shortening adult insect lifespan. Manipulation experiments have demonstrated that the severity of Wolbachia-induced effects on the host is determined by a combination of host genotype, Wolbachia strain, host tissue localization, and interaction with the environment. The recent completion of the whole genome sequence of Wolbachia pipientis wMel strain indicates that it is likely to use a type IV secretion system to establish and maintain infection in its host. Moreover, an unusual abundance of genes encoding proteins with eukaryotic-like ankyrin repeat domains suggest a function in the various described phenotypic effects in hosts.

Wolbachia infection in the Coffee Berry Borer (Coleoptera: Scolytidae)

A nested polymerase chain reaction protocol yielded positive detection of the maternally inherited cytoplasmic proteobacterium Wolbachia in total genomic DNA from coffee berry borers collected in Benin, Brazil, Colombia, Ecuador, El Salvador, Honduras, India, Kenya, Mexico, Nicaragua, and Uganda. Wolbachia was not detected in specimens from Cameroon, the Dominican Republic, Indonesia, Jamaica, and Peru. Amplified bands from India and Brazil were cloned and sequenced. The 438-bp sequence clearly conformed to Wolbachia group B and was nearly identical to that of Ephestia kuehniella. The possible implications of Wolbachia infection in the coffee berry borer are discussed.

Wolbachia infection and expression of cytoplasmic incompatibility in Armigeres subalbatus (Diptera: Culicidae)

Polymerase chain reaction screening revealed that Armigeres subalbatus (Coquillett), a vector of filariasis, was infected with the intracellular bacteria Wolbachia. Laboratory crosses between infected males and uninfected females resulted in less than half the number of offspring than control crosses between uninfected individuals when young (2- to 3-d-old) males were used in the cross. However, incompatibility was lost when old (14- to 17-d-old) males were used. Field-collected females did not show detectable cytoplasmic incompatibility, and this may be because of the age at which males mate in the field. We used head pigment fluorescence levels to age field males collected from mating swarms, and found that 25-63% of swarming males were older than 13 d. Male age may be one factor influencing the observed low levels of cytoplasmic incompatibility detected in the field.

A stable triple Wolbachia infection in Drosophila with nearly additive incompatibility effects

Drosophila simulans strains infected with three different Wolbachia strains were generated by experimental injection of a third symbiont into a naturally double-infected strain. This transfer led to a substantial increase in total Wolbachia density in the host strain. Each of the three symbionts was stably transmitted in the presence of the other two. Triple-infected males were incompatible with double-infected females. No evidence was obtained for interference between modification effects of the different Wolbachia strains in males. Some incompatibility was observed between triple-infected males and females. However, this incompatibility reaction is not a specific property of triple-infected flies, because it was also observed in double-infected strains.