Improved attachment and parasitism of <Emphasis Type="Italic">Trichoderma</Emphasis> on <Emphasis Type="Italic">Meloidogyne javanica</Emphasis> in vitro

Monoclonal and polyclonal antibodies that bind to eggs and/or second-stage juveniles of the nematode Meloidogyne javanica were tested for their effects on the parasitic interactions between this nematode and the fungus Trichoderma. Parasitism of Trichoderma asperellum-203 and Trichoderma atroviride on nematode egg masses, eggs and juveniles was enhanced when antibodies were incorporated into in vitro parasitism bioassays. Parasitism on separated eggs (without gelatinous matrix) and their hatched juveniles was also improved, compared to controls without antibodies that did not attach fungal conidia. Improved parasitism could be due to bilateral binding of the antibodies to the nematodes and conidia, enabling better conidial attachment to the nematodes. Enhanced germination of antibody-bound conidia further improved parasitism. Differences were observed among antibodies in their effects on fungal parasitism and their interaction with Trichoderma species. We focused mainly on the egg- and juvenile-binding monoclonal antibody MISC that exhibited a stronger reaction with T. asperellum-203 than with T. atroviride. Pretreatment of this antibody with fucose inhibited its binding to nematodes and conidial attachment to nematodes, as well as conidial agglutination in the presence of the antibody. Antibody binding to juveniles affected their movement and viability, especially gelatinous matrix-originated juveniles. The fucose-specific lectin Ulex europaeus-I enhanced conidial attachment to nematode life-stages, and conidial agglutination occurred in its presence. These phenomena were inhibited by preincubating lectin with fucose. Our results suggest that carbohydrate residues, such as fucose, on the surface of the nematode and fungal conidia are involved in the antibody- and lectin-mediated improved parasitism.     

Evaluation of Trichoderma spp. as a Biocontrol Agent Against Soilborne Fungi and Plant-parasitic Nematodes in Israel

The optimal conditions required to market Trichoderma as a biocontrol agent against soilborne fungi and nematodes are discussed. These include a proper formulation, an efficient delivery system, and alternative methods for Trichoderma's application.The implementation of Trichoderma in integrated pest management (IPM) can be achieved using a soil treatment which combines reduced amounts of biocides/fungicides and the Trichoderma preparation. Biocontrol activity can be increased by combining two (or more) types of biocontrol agents. Moreover, the construction of a genetically modified Trichoderma can lead to the improvement of certain traits which are absent or not highly expressed in the native microorganism isolated from its natural habitat.Different Trichoderma harzianum and T. lignorum isolates were tested for their nematicidal activity against the root-knot nematode Meloidogyne javanica. In short-term experiments, improved growth of nematode-infected plants and decreases in the root-galling index and the number of eggs per gram of root were achieved when nematode-infested soils were pre- exposed to the T. harzianum preparations. A long-term experiment resulted in improved growth and higher yield of nematode-infected plants, but no significant change in the galling index, either by pre-exposure of the fungus to the soil or by enrichment in the root-ball.As biocontrol is an integral part of the IPM philosophy, judicious use of Trichoderma against soilborne pathogens, when demonstrated to be consistently effective, practical and economic, can serve as a model for the introduction and implementation of other biocontrol means into IPM.     

Renal abscess in a dog with transient diabetes mellitus

A nine-year-old, intact female dalmatian with diabetes mellitus and a renal abscess is described. The renal abscess was treated surgically by nephrectomy, and the diabetes mellitus resolved with ovariohysterectomy. Abdominal ultrasound and ultrasound-guided aspiration of the abscess were helpful in establishing a diagnosis. To the authors' knowledge, this is the first report of a renal abscess in a dog with diabetes mellitus.     

Isospora carliae sp. n. (Apicomplexa: Eimeriidae) from the skink Carlia rhomboidalis (Peters) from Daintree Forest, North Queensland, Australia: description and fine-structural account of endogenous development

Isospora carliae sp. n. is described from the blue-throated rainbow skink Carlia rhomboidalis (Peters), from Daintree Forest, North Queensland, Australia. Oocysts are ellipsoidal, 16.8-21.0 x 12.6-15.4 microm in size, with their two sporocysts, 9.0-14.0 x 7.0-9.24 microm in size, positioned along the wide axis. Sporozoites contain a distinct refractile body and are accompanied by a residuum. All endogenous development occurs within the host-cell nucleus. Nuclei are sometimes invaded by several merozoites, but only infections by a single parasite persist. Nuclei lodging meronts, mature microgamonts and premature macrogamonts have an elongate shape. Some meronts exhibit a membrane-bound cytoplasmic inclusion that contains many micronemes.     

Parasitism of <Emphasis Type="Italic">Trichoderma</Emphasis> on <Emphasis Type="Italic">Meloidogyne javanica</Emphasis> and role of the gelatinous matrix

Trichoderma (T. asperellum-203, 44 and GH11; T. atroviride-IMI 206040 and T. harzianum-248) parasitism on Meloidogyne javanica life stages was examined in vitro. Conidium attachment and parasitism differed beween the fungi. Egg masses, their derived eggs and second-stage juveniles (J2) were parasitized by Trichoderma asperellum-203, 44, and T. atroviride following conidium attachment. Trichoderma asperellum-GH11 attached to the nematodes but exhibited reduced penetration, whereas growth of T. harzianum-248 attached to egg masses was inhibited. Only a few conidia of the different fungi were attached to eggs and J2s without gelatinous matrix; the eggs were penetrated and parasitized by few hyphae, while J2s were rarely parasitized by the fungi. The gelatinous matrix specifically induced J2 immobilization by T. asperellum-203, 44 and T. atroviride metabolites that immobilized the J2s. A constitutive-GFP-expressing T. asperellum-203 construct was used to visualize fungal penetration of the nematodes. Scanning electron microscopy revealed the formation of coiling and appressorium-like structures upon attachment and parasitism by T. asperellum-203 and T. atroviride. Gelatinous matrix agglutinated T. asperellum-203 and T. atroviride conidia, a process that was Ca²⁺-dependent. Conidium agglutination was inhibited by carbohydrates, including fucose, as was conidium attachment to the nematodes. All but T. harzianum could grow on the gelatinous matrix, which enhanced conidium germination. A biomimetic system based on gelatinous-matrix-coated nylon fibers demonstrated the role of the matrix in parasitism: T. asperellum-203 and T. atroviride conidia attached specifically to the gelatinous-matrix-coated fibers and parasitic growth patterns, such as coiling, branching and appressoria-like structures, were induced in both fungi, similarly to those observed during nematode parasitism. All Trichoderma isolates exhibited nematode biocontrol activity in pot experiments with tomato plants. Parasitic interactions were demonstrated in planta: females and egg masses dissected from tomato roots grown in T. asperellum-203-treated soil were examined and found to be parasitized by the fungus. This study demonstrates biocontrol activities of Trichoderma isolates and their parasitic capabilities on M. javanica, elucidating the importance of the gelatinous matrix in the fungal parasitism.     

Eight New Peptaibols from Sponge-Associated Trichoderma atroviride

Eight new and four known peptaibols were isolated from a strain of the fungus, Trichoderma atroviride (NF16), which was cultured from an Axinellid sponge collected from the East Mediterranean coast of Israel. The structures of the pure compounds were determined using HRMS, MS/MS and one- and two-dimensional NMR measurements. The isolated compounds belong to the trichorzianines, a family of 19-residue linear hydrophobic peptides containing a high proportion of α-aminoisobutyric acid (Aib), an acetylated N-terminus and a C-terminal amino alcohol. These new peptaibols exhibited antimicrobial activity against environmental bacteria isolated from the Mediterranean coast of Israel.     

Shifting the natural spring–summer breeding season of the Australian freshwater crayfish Cherax quadricarinatus into the winter by environmental manipulations

Ninety sexually mature Cherax quadricarinatus females were exposed to various combinations of photoperiod and temperature for 2 months during the summer. Females were randomly assigned to either “winter” “semi-winter” or “summer” simulation treatments. In the “winter” treatment, crayfish were exposed to a simulated winter photoperiod (gradual decrease from 14L:10D to 10L:14D, 4 weeks at short day length followed by gradual increase to 14L:10D) and temperature (gradual decrease from 27 to 15 °C, held for 4 weeks, and then gradual increase to 27 °C). In the “semi-winter” treatment, crayfish were exposed to a simulated winter photoperiod and a summer temperature (27–29 °C). In the “summer” treatment, the crayfish were exposed to summer water temperatures and a photoperiod of 14L:10D. Following the 2 months of conditioning, the females were stocked for 7 months in small groups with males under environmental conditions similar to those of the “summer” treatment. All females were individually tagged and molting and spawning events were monitored. Females exposed to “semi-winter” conditioning in the summer, demonstrated a threefold increase in the rate of first spawning during the winter (December–February) compared with the other females. Crayfish breeders can easily implement these findings since shifting the breeding season into the winter only requires shortening of the photoperiod in the summer. The stocking of ponds in the spring with large nursed juveniles that hatched from eggs spawned in the winter, would allow the attainment of market size at the end of the limited growout season in temperate zones.     

Epitheliocystis infection in wild and cultured sea bream (Sparus aurata, Sparidae) and grey mullets (Liza ramada, Mugilidae)

Heavy infection with epitheliocystis on gills caused mortality in hatchery-born Sparus aurata. Histopathological findings showed extensive proliferation of the gill epithelium around the epitheliocystis capsule. Such proliferation was not observed in benign infections. Epizootic infections thus occurred only in hatchery-born S. aurata. In S. aurata and mullets reared from natural stocks, infection remained benign and sporadic. Epitheliocystis infections were also found in juvenile mullets from the east Mediterranean waters as well as from the Gulf of Elat, Red Sea.