Search for hydrophilic marine fungal metabolites: a rational approach for their production and extraction in a bioactivity screening context

In the search for bioactive natural products, our lab screens hydrophobic extracts from marine fungal strains. While hydrophilic active substances were recently identified from marine macro-organisms, there was a lack of reported metabolites in the marine fungi area. As such, we decided to develop a general procedure for screening of hydrophobic metabolites. The aim of this study was to compare different processes of fermentation and extraction, using six representative marine fungal strains, in order to define the optimized method for production. The parameters studied were (a) which polar solvent to select, (b) which fermentation method to choose between solid and liquid cultures, (c) which raw material, the mycelium or its medium, to extract and (d) which extraction process to apply. The biochemical analysis and biological evaluations of obtained extracts led to the conclusion that the culture of marine fungi by agar surface fermentation followed by the separate extraction of the mycelium and its medium by a cryo-crushing and an enzymatic digestion with agarase, respectively, was the best procedure when screening for hydrophilic bioactive metabolites. During this development, several bioactivities were detected, confirming the potential of hydrophilic crude extracts in the search for bioactive natural products.     

Orchardgrass and perennial ryegrass with applied nitrogen and in mixtures with legumes

High-yielding grass-legume mixtures play an important role in forage-animal systems but finding compatible and adapted species can sometimes be difficult. The objective of this study was to examine productivity of perennial ryegrass and orchardgrass in pure stands with N and in mixtures with legumes.
Broadcast sowings were made on conventionally prepared seedbeds in August 1979 and May 1980 on a Hagerstown silt loam soil (fine, mixed mesic Typic Hapludalf). When sown alone orchardgrass cv. Pennlate and perennial ryegrass cv. Reveille received rates of N ranging from 0 to 448 kg ha⁻¹ a⁻¹. The grasses were also grown in mixtures with alfalfa cv. Arc, red clover cv. Arlington, or birds foot trefoil cv. Viking. The same legumes were sown alone. Four legume seed rates were used in both the pure legume and mixed stands.
Orchardgrass-N swards were more productive than ryegrass-N swards over 3 years. More dry matter (DM) was harvested from pure stands of     

Prevalence of Potentially Pathogenic Bacteria as Genital Pathogens in Dairy Cattle

Bacteria on the genital mucosa have been studied less in healthy, non-puerperal cows than in cows with puerperal endometritis. We have thus analysed bacteria in swabs from the vagina and cervix of post-puerperal cattle (n = 644). Out of the animals, 6.8% had aborted within the last 12 months, 2.6% and 11.6% showed signs of vaginitis and endometritis, respectively. In 17.2% of cervical swabs pathogenic gram-positive and in 11.5% pathogenic gram-negative bacteria were found. Arcanobacterium pyogenes was isolated from 41.3% of cows with endometritis and from 3.5% without endometritis (p < 0.05). From 12.5% of cows with abortion but from no cow without abortion, Staphylococcus aureus was recovered (p < 0.05). Out of 383 vaginal swabs, 88.3% were positive. In 3.4% of swabs pathogenic gram-positive and in 16.7% pathogenic gram-negative microorganisms were found. The percentage of positive vaginal swabs did not differ between pregnant and non-pregnant animals. In the genital tract, the percentage of swabs positive for normal mucosal bacteria decreased from caudally to cranially (p < 0.05). Pathogenic bacteria were found more often in cervical than in vaginal swabs (p < 0.05). In conclusion, bacteria on the vaginal and cervical mucosa in cattle involve a wide range of species. In animals without endometritis or vaginitis, colonization of the mucosa rather than infection has to be assumed.     

Aetiology of an ulcerative disease in goldfish, Carassius auratus (L.): experimental induction of the disease

Abstract. Infection experiments were conducted to determine the primary aetiology of an ulcerative disease in goldish, Carassius auratus (L.). Goldfish were exposed to atypical Aeromonas salmonicida and A. hydrophila , previousl y isolated from cutaneous ulcers, and to 04 5 μm filtrates of cutaneous ulcers and kidneys from diseased fish. Fish were exposed to each preparation by intraperitoneal, intramuscular or subcutaneous injection and by a method in which a small patch of scales was removed from each side of the fish and the inoculums applied. Most of the fish injected with A. salmonicida died without developing coetaneous ulcers; however, ulcers were induced in five of the ten fish exposed by the scale removal technique. Exposure to ultra filtrates or A. hydrophila did not result in consistent ulcer formation o r death. Additional experiments showed that a 30-min exposure of goldfish, without prior treatment, to water containing 3 × 10⁵ colony forming units (cfu/ml) of A. salnumicida was sufficient to produce cutaneous lesions in nine often fish exposed. Multiple lesions were produced in most fish and A. salmonicida was consistently recovered. Fish exposed by similar waterborne challenge s to 6·2 × 10⁶ cfu/ml of A. hydrophila or to 7·2 × 10⁶ cfu/ml of another lesion isolate identified as a member of the A. hydrophila complex produced no lesions, eve n when scales were removed. The studies demonstrate that atypical A. salmonicida can initiate cutaneous lesion s characteristic of ulcerative disease, while A. hydrophila and an A. hydrophila complex organism cannot.     

Structural Properties of Starch in Bread and Bread Model Systems: Influence of an Antistaling α‐Amylase

The influence of an antistaling α‐amylase on bread crumb and on wheat starch gels was investigated taking into account different levels of structural hierarchy. Bread was prepared by a conventional baking procedure. Starch gels were produced by heating a concentrated starch dispersion in closed molds. Bread and starch gels were characterized by compression tests, light microscopy (LM), differential scanning calorimetry, and X‐ray measurements. The α‐amylase enhanced the initial firmness of starch gels and reduced the firming rate of bread and starch gels on aging. LM revealed that amylose and amylopectin phase‐separated within the starch granules and that freshly baked control bread and starch gels showed weak birefringence which became more intense during aging. Amylase‐containing bread and starch gels exhibited strong birefringence in the amylose rich region of the granules directly after baking which did not significantly increase during aging. The enzyme hindered the retrogradation of amylopectin as detected by differential scanning calorimetry, whereas X‐ray diffraction indicated that the enzyme induced low levels of starch crystallinity which did not change during aging. It is hypothesized that the antistaling effect of the amylase is based on the capacity to partially degrade amylopectin and, by this, to hinder its recrystallization. On the other hand, the enzyme slightly degrades amylose by an endo‐mechanism which, in turn, promotes the rapid formation of a partly crystalline amylose network in fresh bread and hinders amylose rearrangements during aging.