Mixed Herbs Drugs: Inhibitory Effect on Growth of the Endogenous Mycoflora and Aflatoxin Production

Twenty commercial mixed herbal drugs were examined for mycological profile. Aspergillus species were the predominant fungi found in the drugs. Other fungi harboured in the drugs with less frequency were Paecilomyces species, Eurotium species, Monascus species, Acremonium species, Penicillium species, Cladosporium species, Scopulariopsis species, Phialophora species and Fonseceae species. Fungal count was between 1.0 log₁₀ CFU and 2.4 log₁₀ CFU per gram of sample. When the drugs were incubated in 85% humidity at 25°C, fungal colonies grew on only two of the drugs. The mixed herbal drugs were extracted with water and the extracts were used to grow Aspergillus parasiticus. All extracts reduced aflatoxin B₁ and aflatoxin G₁ production by 62–97%. All but two of the extracts reduced aflatoxin B₂ and aflatoxin G₂ production by 39–95%. It can be concluded that the commercial powdered mixed herbal drugs contained low number of endogenous fungi, and these drugs are inhibitory to the growth of its endogenous fungi and aflatoxins production by aflatoxigenic fungi.     

Aflatoxin B<Subscript>1</Subscript> in chilies from the Punjab region,Pakistan

The occurrence of aflatoxin B1 (AFB1) in chilies from Pakistan was determined by using HPLC in work undertaken in Pakistan. Whole (n = 22) and powdered (n = 22) chilies were analyzed. Sixteen (73.0%) and 19 (86.4%) samples of whole and ground chilies, respectively, were contaminated. The mean concentration in powdered chilies (32.20 μg/kg) was higher statistically than in whole chilies (24.69 μg/kg). Concentrations ranged from 0.00 to 89.56 μg/kg for powdered chilies, compared with 0.00–96.3 μg/kg for whole chilies. The limits of detection and quantification were 0.05 μg/kg and 0.53 μg/kg, respectively. The concentrations were high in general and greater than the statutory limit set by the European Union. There is considerable scope for improvements in chili production in Pakistan.     

Qualitative and quantitative characteristics of rotavirus-specific CD8 T cells vary depending on the route of infection

CD8 T-cell response provides an important defense against rotavirus, which infects a variety of systemic locations in addition to the gut. Here we investigated the distribution, phenotype, and function of rotavirus-specific CD8 T cells in multiple organs after rotavirus infection initiated via the intranasal, oral, or intramuscular route. The highest level of virus-specific CD8 T cells was observed in the Peyer's patches of orally infected mice and in the lungs of intranasally infected animals. Very low levels of virus-specific CD8 T cells were detected in peripheral blood or spleen irrespective of the route of infection. Rotavirus-specific CD8 T cells from Peyer's patches of orally infected mice expressed high levels of CCR9, while CXCR6 and LFA-1 expression was associated with virus-specific CD8 T cells in lungs of intranasally infected mice. Oral infection induced the highest proportion of gamma interferon(-) CD107a/b(+) CD8 T cells in Peyer's patches. When equal numbers of rotavirus-specific CD8 T cells were transferred into Rag-1 knockout mice chronically infected with rotavirus, the donor cells derived from Peyer's patches of orally infected mice were more efficient than those derived from lungs of intranasally infected animals in clearing intestinal infection. These results suggest that different routes of infection induce virus-specific CD8 T cells with distinct homing phenotypes and effector functions as well as variable abilities to clear infection.     

The type and yield of lipopolysaccharide from symbiotically deficient rhizobium lipopolysaccharide mutants vary depending on the extraction method

At least 18 lipopolysaccharide (LPS) extraction methods are available, and no single method is universally applicable. Here, the LPSs from four R.etli, one R.leguminosarum bv. trifolii mutant, 24AR, and the R.etli parent strain, CE3, were isolated by hot phenol/water (phi;/W), and phenol/EDTA/triethylamine (phi/EDTA/TEA) extraction. The LPS in various preparations was quantified, analyzed by deoxycholate polyacrylamide gel electrophoresis (DOC-PAGE), and by immunoblotting. These rhizobia normally have two prominent LPS forms: LPS I, which has O-polysaccharide, and LPS II, which has none. The LPS forms obtained depend on the method of extraction and vary depending on the mutant that is extracted. Both methods extract LPS I and LPS II from CE3. The phi/EDTA/TEA, but not the phi/W, method extracts LPS I from mutants CE358 and CE359. Conversely, the phi;/W but not the phi;/EDTA/TEA method extracts CE359 LPS V, an LPS form with a truncated O-polysaccharide. phi/EDTA/TEA extraction of mutant CE406 gives good yields of LPS I and II, while phi/W extraction gives very small amounts of LPS I. The LPS yield from all the strains using phi/EDTA/TEA extraction is fairly consistent (3-fold range), while the yields from phi/W extraction are highly variable (850-fold range). The phi/EDTA/TEA method extracts LPS I and LPS II from mutant 24AR, but the phi/W method partitions LPS II exclusively into the phenol phase, making its recovery difficult. Overall, phi/EDTA/TEA extraction yields more forms of LPS from the mutants and provides a simpler, faster, and less hazardous alternative to phi/W extraction. Nevertheless, it is concluded that careful analysis of any LPS mutant requires the use of more than one extraction method.     

Testosterone and cortisol concentrations vary with reproductive status in wild female red deer

Although hormones are key regulators of many fitness and life history traits, the causes of individual level variation in hormones, particularly in wild systems, remain understudied. Whilst we know that androgen and glucocorticoid levels vary within and among individuals in mammalian populations, how this relates to key reproductive processes such as gestation and lactation, and their effects on a female''s measurable hormone levels are poorly understood in wild systems. Using fecal samples collected from females in a wild red deer population between 2001 and 2013, we explore how fecal androgen (FAM) and cortisol (FCM) metabolite concentrations change with age and season, and how individual differences relate to variation in reproductive state. Both FAM and FCM levels increase toward parturition, although this only affects FCM levels in older females. FCM levels are also higher when females suckle a male rather than a female calf, possibly due to the higher energetic costs of raising a son. This illustrates the importance of accounting for a female''s life history and current reproductive status, as well as temporal variation, when examining individual differences in hormone levels. We discuss these findings in relation to other studies of mammalian systems and in particular to the relatively scarce information on variation in natural levels of hormones in wild populations.     

Vegetative compatibility and phenotypic characterization as a means of determining genetic diversity of Aspergillus flavus isolates

Toxigenic Aspergillus species produce mycotoxins that are carcinogenic, hepatotoxic and teratogenic immunosuppressing agents in both human and animals. Kenya frequently experiences outbreaks of aflatoxicosis with the worst occurring in 2010, which resulted in 215 deaths. We examined the possible reasons for these frequent aflatoxicosis outbreaks in Kenya by studying Aspergillus flavus diversity, phenotypes and mycotoxin profiles across various agricultural regions. Using diagonal transect random sampling, maize kernels were collected from Makueni, Homa Bay, Nandi, and Kisumu counties. Out of 37 isolates, nitrate non-utilizing auxotrophs complementation test revealed 20 vegetative compatibility groups. We designated these groups by the prefix “KVCG”, where “K” represented Kenya and consequently assigned numbers 1–20 based on our findings. KVCG14 and KVCG15 had highest distribution frequency (n = 13; 10.8 %). The distribution of the L-, S- and S-/L-morphotypes across the regions were 57 % (n = 21); 7 % (n = 3) and 36 % (n = 13), respectively. Furthermore, a unique isolate (KSM015) was identified that had characteristics of S-morphotype, but produced both aflatoxins B and G. Coconut agar medium (CAM) assay, TLC and HPLC analyses confirmed the presence or absence of aflatoxins in selected toxigenic and atoxigenic isolates. Diversity index (H′) analyses ranged from 0.11 (Nandi samples) to 0.32 (Kisumu samples). Heterokaryon compatibility ranged from 33 % (for the Makueni samples, n = 3) to 67 % (Nandi samples, n = 6). To our knowledge, this is the first reported findings for A. flavus diversity and distribution in Nandi, Homa Bay and Kisumu counties and may assist current and future researchers in the selection of biocontrol strategies to mitigate aflatoxin contamination as has been researched in Makueni and neighbouring counties.     

Changes in plasma angiotensin-converting enzyme activity and noradrenaline responses to long-term nitric oxide inhibition vary depending on their basal values in chickens

In this study, we investigated the effects of N(omega)-nitro-L-arginine (L-NNA) on arterial blood pressure (BP), plasma noradrenaline (NA) and adrenaline (A) levels and angiotensin-converting enzyme (ACE) activity. L-NNA was applied with tap water (1 mg/ml) from the 3rd to the 8th week of age (group L-NNA1). In Experiment 1, long-term L-NNA application increased BP compared to the control group (group C1) (L-NNA1 = 131.4 +/- 6.3, n = 6; C1 = 82.7 +/- 4.7 mm Hg, n = 7) but decreased plasma noradrenaline and adrenaline levels and ACE activity (NA levels: C1 = 15.5 +/- 0.8, n = 7; L-NNA1 = 8.6 +/- 0.5 ng/ml, n = 7; A levels: C1 = 15.5 +/- 0.8, n = 7; L-NNA1 = 6.0 +/- 0.5 ng/ml, n = 7; ACE activities: C1 = 87.3 +/- 3.1, n = 6; L-NNA1 = 46.2 +/- 1.9 U/l, n = 5). On the other hand, in Experiment 2 (carried out under the same conditions and in age-matched chickens), blood pressure, plasma noradrenaline levels and ACE activity were found to differ in the control group (C2) (BP = 141.4 +/- 15.5 mm Hg, n = 7; NA = 1.1 +/- 0.4 ng/ml, n = 7; ACE = 57.2 +/- 5.3 U/l, n = 7) as compared to C1, while plasma adrenaline levels were similar. In this series, long-term L-NNA application (group L-NNA2) did not change the BP, but surprisingly increased noradrenaline and ACE values (values of L-NNA2: BP = 165.7 +/- 15.6 mm Hg, n = 7; NA = 9.3 +/- 1.3 ng/ml, n = 8; ACE = 149.4 +/- 16 U/l, n = 8) while decreasing plasma adrenaline levels. L-arginine addition to L-NNA treatment completely reversed plasma noradrenaline and ACE activity values. These results indicate the modulatory activity of an L-arginine-NO pathway on adrenaline release as well as on the renin-angiotensin system in chickens.     

Potential pathogenicity of Cladosporium tennuisimum,Phaeoisaria clematidis and Ramichloridium subulatum in a mouse model

In Sri Lanka, rice is the main staple which is mostly processed into parboiled rice. The levels of aflatoxin B₁ (AFB₁) and aflatoxin G₁ (AFG₁) in parboiled and raw milled rice collected from major rice producing areas and rice consuming townships were estimated. In almost all the samples of parboiled rice examined, the AFB₁ and AFG₁ contents were significantly higher than in raw milled rice. The highest AFB₁ content was 185 µg/kg and AFG₁ content 963 g/kg. These samples were collected from a major rice producing/milling district where the mean relative humidity is 78% and mean annual temperature 27 °C which is the highest amongst the rice growing areas in Sri Lanka. Raw rice was either free of aflatoxins or when toxins were detected, they occurred in less than 10% of the samples. The frequency of occurrence of surface fungal flora (Aspergillus/Penicillium) and aflatoxin content in market samples was closely related. Brownish or greenish moldy rice samples with fermented odour contained over 1000 g/kg of AFB₁.     

The change of Cr and Mn concentrations in selected plants in Samsun city center depending on traffic density

Landscape and Ecological Engineering - In recent years, air pollution has increased significantly due to anthropogenic factors and has become a global problem. Heavy metals in air pollution...     

Hydrogen sulfide concentrations in indoor air of thermal springs

This study aimed to assess hydrogen sulfide (H₂S) concentrations in the pool indoor air of hot springs and its link to some physicochemical properties of the springs in Ardabil province. Twenty-two hot springs from different regions were selected and monitored for H₂S concentrations using a portable gas meter. Respective mean concentrations of H₂S in hot springs indoor air were 9.11 ± 11. Ghotour Souei hot springs had the highest concentration of H₂S with a mean concentration of 29.4 ± 7.7 and 25.2 ± 8.16 ppm at the source and general pool areas, respectively. Oxidation-reduction potential and pH of the water were the most important factor influencing H₂S concentrations in the hot springs. H₂S concentrations in indoor air of Ardabil hot springs were noticeably higher than OEL-TWA and OEL-STEL, and therefore may pose important risks for human health on both short-and long-term exposures.     

Canopy throughfall of Picea abies (L.) Karst. as depending on trace gas concentrations

At six sites in central Germany consequences of SO₂, NO_X and O₃ deposition and of acid precipitation on canopy throughfall of sulphate, nitrate, ammonium, organic acids and of metal cations from Norway spruce crowns were investigated in the field. Measured canopy throughfall rates (mmol ion kg^-1 needle dw a^-1 are separated in (i) background ion throughfall rates in clean air and (ii) trace gas-(or acid interception)-dependent throughfall rates at ambient trace gas concentrations. Based on synchronously measured pollution, precipitation and canopy throughfall data, statistical response functions are given, which allow the separate estimation of annual rates of sulphur and nitrogen deposition into spruce canopies if only annual means of SO₂ or NO₂ concentrations in air are known. The specific SO₂ deposition rate of (0.841±0.214) mmol S kg^-1 needle dw a^-1 (nPa SO₂ Pa^-1)^-1 is 2.3 times higher than the specific stomatal SO₂ uptake. The NO₂-dependent nitrogen deposition of (2.464±0.707) mmol N kg^-1 needle dw a^-1 (nPa NO₂ Pa^-1)^-1 is 2.2 times higher than the specific stomatal NO_X (NO₂+NO) uptake. These ratios (2.32.2) are explained by the percentage of annual hours with open needle stomata. The shape of observed epicuticular SO₂ and NO_X deposition curves and of stomatal SO₂ and NO_X uptake curves are congruent. As for stomatal NO_X uptake, there is an apparent compensation point at (5 to 8) nPa NO₂ Pa^-1. There is significant SO₂-dependent canopy throughfall of Ca>K>Al>Mg>Fe in this order of relative importance. NO_X deposition in spruce canopies reduces K⁺ throughfall and it weakly promotes throughfall of Mn²⁺ and Zn²⁺. There was no significant codeposition of sulphate and ammonium and no ion exchange of intercepted H₃O⁺ with nutrient cations at the measured ambient pH values of the precipitation water. In the presence of O₃, throughfall of Mn²⁺ is reduced and throughfall of K⁺, Ca²⁺ and Al³⁺ is enhanced. In the cooperative presence of SO₂, NO₂ and O₃ pollution in the field there is a 1.3-fold increase of the annual K⁺ demand and a 1.5-fold Mg²⁺ demand of spruce canopies relative to the situation in clean air. This trace gas-dependent additional cation demand of spruce canopies corresponds to a needle loss percentage of (23 to 33)% if the additional K⁺ and Mg²⁺ throughfall could not be recycled in spruce ecosystems. Observed canopy thinning ranges from (13 to 26)% at the investigated six spruce stands.Abbreviations A_spec Specific needle surface area per kg needle dry matter (m²kg^-1 needle dw) - A_tot Total needle surface of spruce stands (ha ha^-1) - [gas]_a Ambient trace gas concentration (gas=SO₂; NO₂ or O₃) in air (nPa Pa^-1=ppb) - GP Number of days per annual growth period d a^-1) - IC_H30 ⁺ Acid interception rate (Eq H₃O⁺ kg^-1 needle dw a^-1) - k_o Trace gas-independent ion throughfall rate constant (mmol kg^-1 needle dw a^-1) - k_gas SO₂-,NO₂-or O₃-dependent ion throughfall rate per unit of trace gas pollution (mmol kg^-1 needle dw a^-1 (nPa Pa^-1)^-1) - k_H30 Specific H₃O⁺/Me⁺ exchange ratio (mol mol^-1) - L_o Background throughfall rate at [gas]_a=0 (mmol kg^-1 needle dw a^-1) - L_ion Canopy throughfall rate of ions (mmol kg^-1 needle dw a^-1) - L'_ion Trace gas dependent ion throughfall (mEq kg^-1 needle dw a^-1 (nPa Pa^-1)^-1) - LAI Leaf area index of the canopy (m² projected needle surface m^-2 ground) - Me⁺ Equivalents of metal cations (Eq) - N Stock of needles of spuce stands in the field (kg needle dw ha^-1) - P_% Percentage of needle loss relative to a healthy reference (%) - r Pearson correlation coefficient (no dimension) - R COO^--Sum of all organic anion equivalents Cat⁺ - An^- (Eq kg^-1 needle dw a^-1) - An^- Sum of all measured inorganic anion equivalents (Eq kg^-1 needle dw a^-1) - Cat⁺ Sum of all measured inorganic cation equivalents (Eq kg^-1 needle dw a^-1)     

Stressful colours: corticosterone concentrations in a free-living songbird vary with the spectral composition of experimental illumination

Organisms have evolved under natural daily light/dark cycles for millions of years. These cycles have been disturbed as night-time darkness is increasingly replaced by artificial illumination. Investigating the physiological consequences of free-living organisms in artificially lit environments is crucial to determine whether nocturnal lighting disrupts circadian rhythms, changes behaviour, reduces fitness and ultimately affects population numbers. We make use of a unique, large-scale network of replicated field sites which were experimentally illuminated at night using lampposts emanating either red, green, white or no light to test effect on stress hormone concentrations (corticosterone) in a songbird, the great tit (Parus major). Adults nesting in white-light transects had higher corticosterone concentrations than in the other treatments. We also found a significant interaction between distance to the closest lamppost and treatment type: individuals in red light had higher corticosterone levels when they nested closer to the lamppost than individuals nesting farther away, a decline not observed in the green or dark treatment. Individuals with high corticosterone levels had fewer fledglings, irrespective of treatment. These results show that artificial light can induce changes in individual hormonal phenotype. As these effects vary considerably with light spectrum, it opens the possibility to mitigate these effects by selecting street lighting of specific spectra.     

Effect of citric acid on aflatoxin degradation and on functional and textural properties of extruded sorghum

Sorghum contaminated with a total concentration of aflatoxin B₁ (AFB₁) and aflatoxin B₂ (AFB₂) at 140 ± 7.3 ng/g was extrusion-cooked in a single screw extruder. The temperature profile in the barrel sections of the extruder was 80–150–200 °C. The flour moisture content (M.C.) was adjusted at 200, 250 and 300 g/kg by means of aqueous citric acid at concentrations of 0, 0.5, 1, 2, 4 and 8N. The barrel temperature profile, in combination with the M.C. and the citric acid concentration, significantly affected the extent of aflatoxin reduction in the extruded sorghum. The recovered aflatoxin decreased with an increase in M.C. and acid concentration. The maximum percentage of aflatoxins degraded from cooking the milled sorghum ranged from 17 to 92%. Even when using a severe extrusion condition combined with high citric acid concentrations, acceptable product colour, viscosity, functional and textural properties were obtained.     

Selectivity and grazing impact of microzooplankton on phytoplankton in two subtropical semi-enclosed bays with different chlorophyll concentrations

Microzooplankton grazing rates were compared between two sites (S1 and S2) in the coastal seas of eastern Hong Kong with similar physio-chemical parameters, but different chlorophyll concentrations. During the period from March 2007 to January 2008, six sets of dilution experiments, combined with high performance liquid chromatography and phytoplankton size fractionation (< 200 μm, < 20 μm and < 5 μm), were carried out to study the microzooplankton grazing rate on phytoplankton of different taxonomic groups and sizes. Although total chlorophyll a concentrations were much higher in S1 (4.98-18.42 μg l^− 1) than in S2 (0.29-1.68 μg l^− 1), size composition of phytoplankton was relatively similar between the two sites. Measured as chlorophyll a, phytoplankton growth rates (− 0.84-1.91 d^− 1 in S1; 0.03-2.85 d^− 1 in S2) and microzooplankton grazing rates (0.00-2.26 d^− 1 in S1; 0.00-1.49 d^− 1 in S2) for all three size fractions were similar between the two bays. Phytoplankton growth rates and microzooplankton grazing rates measured as other pigments for phytoplankton of different size fractions did not show strong variations. Microzooplankton grazing impact, expressed as the ratio of microzooplankton grazing rate to phytoplankton growth rate, was generally higher in S1 than in S2, although the difference was not statistically significant. High microzooplankton grazing impact on alloxanthin (1.00-45.85) suggested strong selection toward cryptophytes. Our results provided no evidence for size selective grazing on phytoplankton by microzooplankton.