Theonellamides J and K and 5-cis-Apoa-theopalauamide,Bicyclic Glycopeptides of the Red Sea Sponge Theonella swinhoei

Theonella swinhoei is a fairly common inhabitant of reefs throughout the Indian and Pacific Oceans. Metabolomic analyses of samples of T. swinhoei collected in different depths in the Gulf of Aqaba revealed two chemotypes differing in the profiles of the theonellamides they produce, some of which seem to be unknown. Driven by this finding, we examined a sample of T. swinhoei collected more than 40 years ago in the southern part of the Gulf of Aqaba. Large-scale extract of this sample yielded four theonellamides, the known theopalauamide (4), as the major component, and three new metabolites, theonellamide J (1), 5-cis-Apoa-theopalauamide (2), and theonellamide K (3), as the minor components. The planar structure of these complex cyclic glycopeptides was elucidated by combination of 1D and 2D NMR techniques and HRESIMS. The absolute configuration of the amino acids was established by Marfey’s and advanced Marfey’s methods, and the absolute configuration of its galactose unit using “Tanaka’s method” for monosaccharides. The biological activity of the pure compounds was tested for antibacterial activity and for cytotoxicity to HTC-116 cell line. The compounds presented significant cytotoxicity against the HTC-116 cell line, illuminating the importance of the Apoa subunit for the activity.     

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.     

A selective medium for improving quantitative isolation ofTrichoderma spp. from soil

ATrichoderma-selective agar medium (TSM) was developed for quantitative isolation ofTrichoderma spp. from soil. Selectivity was obtained by using chloramphenicol as a bacterial inhibitor, and pentachloronitrobenzene, p-dimethylaminobenzenediazo sodium sulfonate and rose-bengal as selective fungal inhibitors. The TSM also contains a low concentration of glucose which still allows relatively rapid growth and sporulation ofTrichoderma, enabling convenient and rapid identification ofTrichoderma colonies. All the 15Trichoderma isolates tested formed colonies and grew well on this medium. Recovery ofTrichoderma from artificially inoculated soils was high and was not affected by soil type or by other microorganisms. A positive correlation was observed betweenTrichoderma added to soil and counts ofTrichoderma colonies on TSM plates. When combined with a soil pellet sampler, the selective medium was also used successfully for recovery of the indigenousTrichoderma population of natural soils.     

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.     

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.     

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.     

Control of Rhizoctonia solani fruit rot of tomatoes by Trichoderma harzianum Rifai

Application of Trichoderma harzianum, to soil or by coating tomato fruits, reduced Rhizoctonia solani fruit rot by up to 43% and 85%, respectively, under laboratory conditions. When mixed with naturally infested soil, Trichoderma reduced R. solani inoculum potential by 86% in field trials. It also significantly reduced fruit rot by 27–51%.     

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.