Forensic identification of the guitarfish species Rhinobatos horkelli, R. percellens and Zapteryx brevirostris using multiplex-PCR

Rhinobatos percellens, Rhinobatos horkelli and Zapteryx brevirostris, known as guitarfish, are commonly found inshore and caught by trawl nets. Recently, these species have been suffering sharp population declines. One of the main impediments to the development of conservation plans is the lack of information related to each species catch. Gathering this information is made more difficult by the fishermen's practice of removing parts of the animals before landing, as well as the morphological similarities among the three species. To address these issues, this work presents multiplex-PCR protocols that were developed to distinguish among these species using their genetic characteristics.     

Post-impoundment changes in the fish fauna of Lake Itezhi-tezhi, Zambia

The fish fauna of the newly inundated Lake Itezhi-tezhi, Zambia was observed between 1980 and 1985. Marked changes in the community structure were identified. The most obvious were a decline in species diversity and a shift in species composition from acommunity with a preponderance of Alesles lateralis (Boulenger) (Characidae), to one dominated by cichlids. These changes were primarily due to members of the families Cyprinidae, Mormyridae and Schilbeidae being unable to adapt to the new environmental conditions, such as spawning and feeding grounds, and their inability to cope with fluctuating water levels in the lake. It was suggested that a sound fishery data collection system be implemented to provide reliable information on which to base management policy.     

A capillary zone electrophoresis method for the quantitative determination of hypoxoside in commercial formulations of African potato (Hypoxis hemerocallidea)

Hypoxoside is a norlignan diglucoside present in the corms of African potato, Hypoxis hemerocallidea, used as a popular African traditional medicine for its nutritional and immune boosting properties. A specific analytical method employing capillary zone electrophoresis has been developed and validated for the quantitative determination of this analyte. Sulfafurazole was used as internal standard, and electrophoretic separation of both analytes could be achieved within 12 min. Linearity of the method was established within the range 5-60 microg/mL and provided a high degree of accuracy (100 +/- 3%). The recovery of the method was found to be 100 +/- 5% and the RSDs of the intra- and inter-day precision were better than 5.19 and 2.52%, respectively. The limits of detection and quantification were calculated to be 0.5 and 2 microg/mL, respectively. The described method was used for the analysis and quality control of two commercially available products containing African potato. The method can also be used to determine product stability since it could separate the hypoxoside peak from its degraded products obtained from degradation studies.     

A commercial iron fertilizer increases the survival of Fusarium oxysporum f. sp. orthoceras propagules in a wheat flour-kaolin formulation

Fusarium oxysporum f.sp. orthoceras, a potential biocontrol agent against the parasitic weed Orobanche cumana Wallr., has proven to be efficacous under greenhouse conditions when formulated as wheat-kaolin granules. To help minimize the loss of viable fungal propagules during the formulation process, the addition of a commercial fertilizer containing the iron chelate of EDDHA to the formulation was investigated. The proportion of surviving propagules was significantly increased after adding the fertilizer. However, growing conidia in fertilizer-amended liquid medium did not reduce losses in viability during formulation. The efficacy of the formulated fungus and the storability at room temperature for the first 3 months was not affected by the iron fertilizer. The protective effect could neither be obtained with the chelator EDDHA alone nor with FeEDTA or FeSO_4.     

Evaluation of an experimental product based on Bacillus thuringiensis sorovar. israelensis against Aedes aegypti larvae (Diptera:Culicidae)

The larvicidal activity of an experimental formulation of Bacillus thuringiensis israelensis (Bti) against Aedes aegypti larvae was evaluated under laboratory and simulated field conditions (SFC). Samples of technical powder (TP) were assayed to establish the LC₅₀ and the potency of the product. The larvicidal activity of the TP and the tablet (T) were evaluated under SFC to assess the efficacy and the residual activity, measured against Ae. aegypti larvae. Either a T or 250 mg of TP were added to 50 L of water in plastic containers. Containers were exposed to sunlight or kept in the shade. Results showed a LC₅₀ of 0.26 mg/L and a potency of 750 ITU/mg. In spite of differences in the toxicity amongst TP and T samples, all of them killed 98–100% of the larvae and the mortality remained high for six months, in the shade. The replacement of 20% or 60% of the water volume did not affect the activity of the product. Seasonal differences influenced the persistence of the product in containers exposed to sunlight. Both formulations showed an excellent performance, especially when kept in the shade. The Bti tablet evaluated in this study is potentially very useful in programs to control dengue vectors.     

Interaction between Shaoyao–Gancao–Tang and a laxative with respect to alteration of paeoniflorin metabolism by intestinal bacteria in rats

Shaoyao-Gancao-Tang (SGT), a traditional Chinese herbal medicine (Kampo formulation) containing Shaoyao (Paeoniae Radix) and Gancao (Glycyrrhizae Radix), is co-administered with laxative sodium picosulfate as a premedication for relieving the pain accompanying colonoscopy. Paeoniflorin (PF), an active glycoside of SGT, is metabolized into the antispasmodic agent paeonimetabolin-I (PM-I) by intestinal bacteria after oral administration. The objective of the present study was to investigate whether the co-administered laxative (sodium picosulfate) influences the metabolism of PF to PM-I by intestinal bacteria. We found that the PF-metabolizing activity of intestinal bacteria in rat feces was significantly reduced to approximately 34% of initial levels by a single sodium picosulfate pretreatment and took approximately 6 days to recover. Repeated administration of SGT after the sodium picosulfate pretreatment significantly shortened the recovery period to around 2 days. Similar results were also observed for plasma PM-I concentration. Since PM-I has muscle relaxant activity, the present results suggest that repetitive administration of SGT after sodium picosulfate pretreatment might be useful to relieve the pain associated with colonoscopy.     

Development of a bladder instillation of the indoloquinone anticancer agent EO-9 using tert-butyl alcohol as lyophilization vehicle

The purpose of this research was to develop a stable bladder instillation of EO-9 for the treatment of superficial bladder cancer. First, stability and dissolution studies were performed. Subsequently, the freeze-drying process was optimized by determination of the freeze-drying characteristics of the selected cosolvent/water system and differential scanning calorimetry analysis of the formulation solution. Furthermore, the influence of the freeze-drying process on crystallinity and morphology of the freeze-dried product was determined with x-ray diffraction analysis and scanning electron microscopy, respectively. Subsequently, a reconstitution solution was developed. This study revealed that tert-butyl alcohol (TBA) can be used to both dramatically improve the solubility and stability of EO-9 and to shorten the freeze-drying cycle by increasing the sublimation rate. During freeze drying, 3 TBA crystals were found: TBA hydrate-ice crystals, crystals of TBA hydrate, and a third crystal, probably composed of TBA hydrate crystals containing ≈90% to 95% TBA. Furthermore, it was shown that crystallization of TBA hydrate was inhibited in the presence of both sodium bicarbonate (NaHCO₃) and mannitol. Addition of an annealing step resulted in a minor increase in the crystallinity of the freeze-dried product and formation of the δ-polymorph of mannitol. A stable bladder instillation was obtained after reconstitution of the freeze-dried product (containing 8 mg of EO-9, 20 mg of NaHCO₃, and 50 mg of mannitol per vial) to 20 mL with a reconstitution solution composed of propylene glycol/water for injection (WfI)/NaHCO₃/sodium edetate 60%/40%/2%/0.02% vol/vol/wt/wt, followed by dilution with Wfl to a final volume of 40 mL. Published: August 3, 2007     

Using particle size analysis to determine the hydrophobicity and suspension of fungal conidia with particular relevance to formulation of biopesticide

Fungal formulations are vital for effective biopesticide development. Good formulations help to optimise field efficacy, while poor formulations result in product failure. This study aimed to produce a hydrophobicity test that would be appropriate for fungal conidia produced to a commercial quality and determine relative hydrophobicity of fungi from four different genera by using laser diffraction. A particle size analyser was used to determine the hydrophobicity of: three Metarhizium acridum samples, M. anisopliae, Beauveria bassiana, Trichoderma stromaticum, T. harzianum, T. viride and Alternaria eichhorniae conidia by suspending the conidia in three different liquids: Shellsol T (a mineral oil), water and 0.05 % Tween 80. Hydrophobicity was determined by the size of the particles formed in each of the liquids. All the Metarhizium samples were the most hydrophobic followed by B. bassiana and A. eichhorniae. The Trichoderma samples were the least hydrophobic. As a comparison, a phase exclusion assay and a salt-mediated aggregation and sedimentation (SAS) test were performed. It was not possible to get a reliable reading for the B. bassiana, A. eichhorniae and T. viride samples using the phase exclusion assay. The addition of salt in the SAS test did not affect the rate of sedimentation. It was hypothesised that conidia size affected the results of the SAS test that made A. eichhorniae the most hydrophobic conidia. Particle size analysis (PSA) was a more accurate test for comparing fungi from difference genera compared to the SAS test and phase exclusion assay. PSA was also used to test three emulsions and demonstrated that different formulations had an effect on particle size.     

Comparison of Water,Oils and Emulsifiable Adjuvant Oils as Formulating Agents for <Emphasis Type="Italic">Metarhizium Anisopliae</Emphasis> for Use in Control of <Emphasis Type="Italic">Boophilus Microplus</Emphasis>

Studies were conducted to identify oil-based formulating agents (paraffinic oil, palm oil and emulsifiable adjuvant oils (EAOs)) for Metarhizium anisopliae that were superior to water with simple surfactants using a germination test and a bioassay against Boophilus microplus. Germination of conidia in all formulations, except 10% coconut EAO, produced more than 68% germination at 24 h and nearly 100% at 48 h. Coconut oil (average survival time (AST)=4.6±0.28 days) and 10% liquid paraffin EAO (AST=4.4±0.15 days) enhanced the pathogenicity of M. anisopliae to B. microplus relative to water (AST=8.4±0.42 days). M. anisopliae in 10% liquid paraffin EAO was the most effective formulation having a moderately high germination after 24 h and a low AST as well as a high AST in the control. In the second experiment, germination of conidia in 2% liquid paraffin EAO and 2% Cropspray was higher than in 2% Codacide oil at 24 h, however, all treatments reached 100% germination after 48 h. The ASTs of the EAO based M. anisopliae formulations (Average AST=6.4±0.54 days) were similar but lower that the ASTs of the controls (Average AST=9.6±0.28 days).