Incubation Method Research Articles

Spreader-CDR system for efficient, reproducible, and frugal western blot.

Efficient antibody incubation is a vital step for successful western blot. During the incubation, a thin antibody-depleted layer is created around the blotting membrane, which limits antibody binding. Although the conventional batch shaking method is ineffective against it, this layer can be easily disrupted by cyclic draining and replenishing (CDR) of the antibody solution during membrane incubation. Previously, we introduced a closed and rotating cylindrical chamber as a tool to implement CDR for western blots (rCDR). A new open bucket-style chamber was devised for easier operation and the possibility of process automation. Instead of rotation as in rCDR, rocking it back and forth achieved the CDR antibody incubation (R-CDR). The chamber was then equipped with a spreader-rod to facilitate the uniform movement of the antibody solution across the membrane surface. Hence, it was named spreader CDR (S-CDR). Compared to the batch incubation method, both the S-CDR and R-CDR devices produced significantly enhanced signals and developed faster results. There were several additional benefits of using the spreader-rod, which included uniform antibody binding across the membrane, reduced usage of antibodies, and the ability to recover results even from mishandled, creased membranes. The S-CDR device ensures better blots and can be easily implemented in existing western blot protocols.

Metabolite fingerprinting, in vitro antimicrobial and antioxidant activities and in-silico docking in Alloteropsis cimicina and its endophytic fungus Penicillium pinophilum.

Endophytic fungal interaction with medicinal plant resulting in the production of bioactive compounds influenced the present study. Endophytic fungus Penicillium pinophilum Hedgc. from Alloteropsis cimicina with high incidence was isolated by incubation methods and characterized by morphological and molecular (ITS rDNA region) methods. Penicillium pinophilum was cultured on PD broth and metabolites of host and endophyte were extracted with ethyl acetate and methanol. Metabolites were assayed for antimicrobial potential by well diffusion and scavenging ability by spectrophotometric and electrochemical methods. Metabolite profiling by Orbitrap High-Resolution Liquid Chromatography-Mass Spectrometry and Fourier-transform infrared spectroscopy and by validation of antimicrobial and antioxidant activities in-silico drug-likeness of spectral compound prediction and molecular docking were performed. Host and P. pinophilum extracts showed strong antimicrobial potential against certain clinical bacterial strains and Fusarium oxysporum. Fungal extracts exhibited higher antioxidant activity than A. cimicina extract. Metabolite profiling indicated 14 and 21 antimicrobial, 10 and 13 antioxidant compounds in A. cimicina and P. pinophilum extracts, respectively. There were eight spectral compounds common to endophyte and host with high binding affinity towards receptors. The present study revealed that P. pinophilum and A. cimicina are natural reservoirs of novel bioactive compounds with antimicrobial and antioxidant properties.

Microbiological Study in Petrol-Spiked Soil.

The pollution of arable lands and water with petroleum-derived products is still a valid problem, mainly due the extensive works aimed to improve their production technology to reduce fuel consumption and protect engines. An example of the upgraded fuels is the BP 98 unleaded petrol with Active technology. A pot experiment was carried out in which Eutric Cambisol soil was polluted with petrol to determine its effect on the microbiological and biochemical properties of this soil. Analyses were carried out to determine soil microbiome composition—with the incubation and metagenomic methods, the activity of seven enzymes, and cocksfoot effect on hydrocarbon degradation. The following indices were determined: colony development index (CD); ecophysiological diversity index (EP); index of cocksfoot effect on soil microorganisms and enzymes (IFG); index of petrol effect on soil microorganisms and enzymes (IFP); index of the resistance of microorganisms, enzymes, and cocksfoot to soil pollution with petrol (RS); Shannon–Weaver’s index of bacterial taxa diversity (H); and Shannon–Weaver’s index of hydrocarbon degradation (IDH). The soil pollution with petrol was found to increase population numbers of bacteria and fungi, and Protebacteria phylum abundance as well as to decrease the abundance of Actinobacteria and Acidobacteria phyla. The cultivation of cocksfoot on the petrol-polluted soil had an especially beneficial effect mainly on the bacteria belonging to the Ramlibacter, Pseudoxanthomonas, Mycoplana, and Sphingobium genera. The least susceptible to the soil pollution with petrol and cocksfoot cultivation were the bacteria of the following genera: Kaistobacter, Rhodoplanes, Bacillus, Streptomyces, Paenibacillus, Phenylobacterium, and Terracoccus. Cocksfoot proved effective in the phytoremediation of petrol-polluted soil, as it accelerated hydrocarbon degradation and increased the genetic diversity of bacteria. It additionally enhanced the activities of soil enzymes.

Quality Dahi Preparation in Automated Controlled Ambient Conditions

This automated controlled system eliminates the need of two separate places for incubation of the curd at higher temperature in the range of 39 to 43°C and then shifting the set curd cups into cold rooms for storage purpose and kept at about 4 to 5°C.The transient cooling process of dahi, set in cups and also in steel containers placed in cold air flow was conducted in this experimental study. Recording of the temperature of dahi-cups and of the supplied air were done at regular intervals until the temperature reached below 4°C to 5°C. The exponential curves of cooling by forced convection process for the dimensionless temperature of curd were obtained for the trials conducted at the different velocities of cooling air flow. It was found that the surface heat transfer coefficient increased and duration of cooling decreased by increasing the air velocities from 0.5 m/s by evaluating the Biot number and surface heat transfer coefficient. In the initial period of cooling this pattern was more effective and reduced for higher velocities of air from 3.5 to 4.5 m/s. The method of incubation and storing dahi-cups at the same place and changing the ambient temperature of the whole environment instead of changing the place of dahi-cups for cooling purpose have been applied in this research work to control the problem of whey-off in set-curd.

Single and combined toxicity of amino-functionalized polystyrene nanoparticles with potassium dichromate and copper sulfate on brine shrimp Artemia franciscana larvae.

The increasing use and disposal of plastics has become a persistent problem in the marine environment, calling for studies that refer to realistic scenarios to understand their effects on biota. Particularly, the understanding about the effects of co-exposure with nanoplastic particles and metals on aquatic organisms is still limited. The present work aimed to investigate the acute toxicity of amino-functionalized polystyrene nanoparticles (PS-NH2; 50 nm) as proxy for nanoplastics on brine shrimp Artemia franciscana larvae under different culture conditions and at different stages of development, as well as the combined effect with two reference toxicants - potassium dichromate (K2Cr2O7) and copper sulfate (CuSO4). Nauplii (instar II or III larval stages) were exposed to different concentrations of PS-NH2 (0.005 to 5 μg mL-1) for up to 48 h, with or without agitation in order to mimic a more realistic environmental scenario. Larval mobility and PS-NH2 accumulation were monitored under microscopy. PS-NH2 alone showed toxicity only at the highest concentration tested (5 μg mL-1) regardless the incubation method used (61.2 + 3.1% and 65.0 + 4.5% with and without agitation, respectively). Moreover, instar III stage was the most sensitive to PS-NH2 exposure (38.2% immobility in 24 h of exposure; 5 μg mL-1). Evidence of PS-NH2 retention in the gastrointestinal tract in a concentration- and time-dependent manner was also obtained. Mixtures of PS-NH2 (0.005 and 5 μg mL-1) with different concentrations of K2Cr2O7 increased the immobilization rate of the larvae after 48 h of exposure, when compared to the K2Cr2O7 alone. Similar results were observed for CuSO4 in the co-exposure conditions at different concentrations. However, exposing nauplii to a mixture of PS-NH2 (0.005 μg mL-1) and CuSO4 decreased immobilization rate, in comparison to the group exposed to CuSO4 alone. The present work highlights the potential risk posed by nanoplastics to zooplanktonic species through their interaction with other toxicants.

Plausible drug interaction between cyclophosphamide and voriconazole via inhibition of CYP2B6

The inhibitory activities of eight cytochrome P450 (CYP) isoenzymes for representative or suspected inhibitors of CYPs, including pesticides, were evaluated simultaneously using an in vitro cocktail incubation method to demonstrate the importance of systematic evaluation of CYP inhibitory risks in drug interaction (DI). Potent inhibition of CYP2B6 was noticeable for some azoles, including voriconazole. When voriconazole and cyclophosphamide were co-administered in mice, cyclophosphamide-induced alopecia and leukopenia were significantly suppressed by approximately 50% with increased blood concentrations of cyclophosphamide. The formation of an active metabolite of cyclophosphamide was suppressed effectively by voriconazole in the mouse liver microsomes. Surveys of adverse event reporting databases in Japan (JADER) and the U.S. (FAERS) showed that the proportional reporting ratios of neutropenia, hemorrhagic cystitis, and alopecia for cyclophosphamide, which is principally activated by CYP2B6 in humans, were mostly reduced, or tended to be reduced when azoles, including voriconazole, were prescribed in combination. It is highly likely that DIs between cyclophosphamide and azoles occur in the clinical setting. This study also suggests that more proper consideration of CYP2B6-mediated DIs is warranted. The combination of the in vitro cocktail method and a survey of adverse event reporting databases was a useful method to comprehensively detect pharmacokinetic DIs.

Effects of stocking rate on the interannual patterns of ecosystem biomass and soil nitrogen mineralization in a meadow steppe of northeast China

Understanding the effects of livestock grazing on the ecosystem biomass and soil nitrogen processes of grassland ecosystems is critical to improving knowledge on the mechanisms underlying grassland degradation and accurately assessing the influence of grazing management on grassland functions. We examined the interannual patterns of ecosystem biomass and soil nitrogen mineralization in response to cattle grazing in a Chinese meadow steppe. The soil core incubation method was employed for soil N transfer estimation, whilst the fumigation extraction method, a modified Baermann funnel method and harvest method were used for various measurements of ecosystem biomass parameters. We found that cattle grazing caused consistent significant increases in soil temperature, irrespective of the stocking rate and year, whereas significant effects on soil moisture and edaphic properties were observed only in individual years and/or at specific stocking rates. Consistent positive effects at moderate stocking rates were observed for aboveground net primary production and soil nematode biomass in all study years. The across stocking rate pattern of N mineralization in response to cattle stocking appeared to be year-specific, although negative effects were found in most cases. In contrast, the interannual pattern of N mineralization was determined principally by the interannual patterns in precipitation and soil moisture and was much less affected by cattle grazing. Soil N mineralization in this meadow steppe was affected by cattle grazing via two major mechanisms, i.e., its effects on the aboveground net primary production (ANPP) and thus the quantity of plant litter input into the soil and its effects on soil temperature and moisture. Overall, our study spanned the longest consecutive years with the broadest range of stocking rates thus far of its kind, which revealed for the first time that the soil nitrogen mineralization pattern with respect to stocking rate was year-specific. Our findings have important implications for adaptive management and sustainable utilization of Chinese grasslands.

A Study of Soil Organic Matter Stability Using Derivatography and Long-Term Incubation Methods

Soil organic matter (SOM) includes many classes of labile compounds available for microbial decomposition or, conversely, stable compounds protected from biodegradation by biological, chemical, and physical stabilization. It is believed that the more thermal energy is spent on the destruction of soil organic matter, the more stable and more resistant for biodegradation it is. We compared the thermal and biological stabilities of organic matter in eleven soil types from deciduous forest, forest-steppe, steppe, and semidesert bioclimatic areas of the European Russia. According to the activation energy (Ea), the highest SOM thermal stability was typical of the ordinary chernozem and meadow vertic soil. The lowest SOM thermal stability was found for gray forest soil; other soil types were characterized by an intermediate resistance towards thermal oxidation. The thermally labile pool (<390–400°C) of organic matter in soils was on the average 41% (32–60%) of the total SOM, while the thermally stable pool (>390–400°C) was on the average 59% (40–68%). The SOM biological stability estimated by the ratio of potentially mineralizable organic matter to that resistant to mineralization (biological stability index) decreased in the following order: ordinary chernozem (Haplic Chernozem (Loamic, Pachic)) > meadow vertic soil (Pellic Vertisol (Gleyic, Humic)) > gray forest soil (Luvic Greyzemic Phaeozem (Loamic)) = meadow chestnut soil (Gleyic Kastanozem (Chromic)) > meadow solonetz (Endosalic Gleyic Solonetz (Loamic, Cutanic)) > alluvial meadow soil (Eutric Fluvisol (Humic, Oxyaquic)). The potentially mineralizable SOM pool in the studied soils was 6–27-fold lower as compared with the pool of thermally labile SOM, and the parameters that characterize SOM thermal stability did not correlate with the biological stability index. Thus, SOM thermal lability is not identical to its biodegradability.

Óleos essenciais sobre a qualidade de sementes de leucena

&lt;span class="fontstyle0"&gt;Leucena (Leucaena leucocephala) is an alternative used for animal feeding in the Northeastern of Brazil due to its resistance to drought and its high nutritional value. They are forest species whose seeds are affected by fungi that cause abnormalities and damage to seedlings, seed deterioration. The objective of this study was to evaluate the efficiency of essential oils in reducing the incidence of fungi associated with L. leucocephala seeds and their interference with physiological quality. The treatments were the following: essential oils of Cymbopogon nardus, Boswellia carteri, Eucalyptus globulus, Helianthus annuus, Vitis vinifera seeds, Eugenia caryophyllata, Melaleuca alternifolia and Rosmarinus officinalis in a concentration of 1 mL · L&lt;/span&gt;&lt;span class="fontstyle0"&gt;-1 &lt;/span&gt;&lt;span class="fontstyle0"&gt;and fungicide, using 100 seeds per treatment.In order to overcome dormancy scarification with sandpaper Nº 100 in the opposite region to the micropile. The sanitary quality of the seeds was made with the incubation method in Petri dishes containing double layer of filter paper moistened with ADE. Seed physiological quality was determined based on seed germination, emergence and vigor tests. The experimental design was completely randomized with ten treatments. Eucalyptus, clove and Melaleuca alternifolia essential oils reduced the incidence percentage of fungi associated with leucena seeds. Melaleuca alternifolia essential oil reduced the physiological quality of leucena seeds.&lt;/span&gt; &lt;br /&gt;&lt;br /&gt;

An integrated field-laboratory investigation of the effects of low oxygen and pH on North Pacific krill (Euphausia pacifica)

Krill are abundant and ecologically important zooplankton that inhabit dynamic environments characterized by strong natural variability, but global ocean change is shifting the range of conditions that they experience. Laboratory tests reveal that krill are sensitive to ocean acidification despite residing in naturally low pH areas, showing the importance of modulating factors for determining their responses. This study combines laboratory manipulations with field observations across a strong natural water chemistry gradient in Puget Sound, Washington, USA to investigate the effects of pH and oxygen on adult female North Pacific krill, Euphausia pacifica. Enzyme activities of the Electron Transport System (ETS) and aminoacyl-tRNA synthetases (AARS) were used as indices of zooplankton metabolism and growth, respectively, and were paired with traditional incubation methods. Acclimation to pH and oxygen conditions in the laboratory did not reveal effects on respiration rate, ETS, or AARS activity of krill. However, field observations showed that respiratory potential, as estimated by ETS activity, decreased with decreasing oxygen, declining 9% (95% confidence interval 2.5–15%) over the range of conditions we observed (3.9–8.1 mg O2 L−1). This reduction would depress the metabolic potential of krill in areas of stressful conditions (concurrent low pH), though krill also displayed a high degree of inter-individual variability. Although differences in age structure suggest different patterns of recruitment between E. pacifica populations in areas with stressful conditions and those without, populations persist at stressful sites. Lower temperature of waters with low oxygen and pH, as well as high food concentrations, may contribute to these populations’ success.

Carbon, nitrogen and phosphorus stoichiometry controls interspecific patterns of leaf litter-derived dissolved organic matter biodegradation in subtropical plantations of China

Leaching of leaf litter is the primary source of dissolved organic matter (DOM) in forest soils. However, the interspecific variations of litter-derived DOM characteristics and biodegradation and their controlling factors remain unclear in subtropical plantations. Using fresh leaf litter of two broadleaf trees (Liquidambar formosana and Schima superba) and two coniferous trees (Pinus massoniana and P. elliottii) in subtropical plantations of China, we assessed the effects of tree species on the amounts and properties of litter-derived DOM with a short-term leaching experiment, and examined the interspecific variation of DOM biodegradation using a 56-day laboratory incubation method. Broadleaf tree litter generally leached higher amounts of dissolved organic carbon (DOC), dissolved total nitrogen (DTN), and dissolved total phosphorus (DTP) than coniferous tree litter. Compared with coniferous trees, broadleaf trees had higher DOM aromaticity and molecular weight, but lower DOC:DTP and DTN:DTP ratios in the litter leachates. Despite greater DOM aromaticity and molecular weight, broadleaf trees had higher litter-derived DOM biodegradation than coniferous trees because of the relatively lower DOC:DTP and DTN:DTP ratios. These results indicate the distinct patterns of litter-derived DOM characteristics and biodegradation between broadleaf and coniferous trees, and also highlight the predominant role of C:N:P stoichiometry in driving the interspecific variation of litter-derived DOM biodegradation in subtropical plantations of China.

Longevity of Bipolaris oryzae on HEERA-2 Hybrid Rice Seed

Seeds of hybrid rice variety Heera-2 was collected from Mymensingh district in Bangladesh. Longevity of Bipolaris oryzae was studied in 2012 to 2016 at Seed Pathology Centre, Bangladesh Agricultural University, Seed processing &amp;preservation center and Seed Pathology Laboratory of Supreme Seed Company Limited. The data collected from the experiments were analyzed for test of significance and compared the treatment means by using DMRT at 5% level of probability following the Statistical tool for Agricultural Research (STAR) 2.1 program.The longevity of B. oryzae was observed till five years by blotter incubation methods, seedling symptom test and growing on test. Both untreated and seeds treated with Vitaflo 200FF (carboxin and thiram) were kept in jute bag and polybag stored for five years under both normal and dehumidified condition. The inocula of B. oryzae on Hybrid Heera 2 seed remained viable up to five years. When the seed samples were stored under dehumidified condition, the highest incidence of B. oryzae was recorded in case of jute bag after five years. After treated with Vitaflo 200FF, there was no any infection of B. oryzae under both storage condition&#x0D; Environ. Sci. &amp; Natural Resources, 12(1&amp;2): 209-213, 2019

Extracellular Vesicles Act as Nano-Transporters of Tyrosine Kinase Inhibitors to Revert Iodine Avidity in Thyroid Cancer.

A new approach for using extracellular vesicles (EVs) to deliver tyrosine kinase inhibitors (TKIs) to enhance iodine avidity in radioactive iodine-refractory thyroid cancer is needed. We isolated and characterized primary human adipose-derived stem cells (ADSCs) and isolated their EVs. The EVs were characterized by transmission electron microscopy, nanoparticle tracking analysis, and western blotting. A new TKI was loaded into the EVs by incubation (37 °C; 10 min) or sonication (18 cycles; 4 s per cycle) with 2 s intervals and a 2 min ice bath every six cycles. TKI loading was confirmed and measured by mass spectrometry. EV uptake into radioactive iodine-refractory thyroid cancer cells (SW1736 cells) was confirmed by microscopy. We treated the SW1736 cells with vehicle, TKI, or TKI-loaded EVs (sonication TKI-loaded EVs [EVsTKI(S)]) and examined the expression of iodide-metabolizing proteins and radioiodine uptake in the SW1736 cells. ADSCs cells showed >99% of typical stem cell markers, such as CD90 and CD105. The EVs displayed a round morphology, had an average size of 211.4 ± 3.83 nm, and were positive for CD81 and Alix and negative for cytochrome c. The mass spectrometry results indicate that the sonication method loaded ~4 times more of the TKI than did the incubation method. The EVsTKI(S) were used for further experiments. Higher expression levels of iodide-metabolizing mRNA and proteins in the EVsTKI(S)-treated SW1736 cells than in TKI-treated SW1736 cells were confirmed. EVsTKI(S) treatment enhanced 125I uptake in the recipient SW1736 cells compared with free-TKI treatment. This is the first study that demonstrated successful delivery of a TKI to radioactive iodine-refractory thyroid cancer cells using EVs as the delivery vehicle. This approach can revert radioiodine-resistant thyroid cancer cells back to radioiodine-sensitive thyroid cancer cells.

Temporal distribution of primary and secondary production estimated from water quality data in the Seto Inland Sea, Japan

Estuaries and coastal areas support highly productive fisheries and aquaculture operations. The high productivity is partly supported by inputs of nutrients from land; consequently, these areas are sensitive and vulnerable to human activities. Excess anthropogenic nutrient input can lead to accelerated phytoplankton growth and associated environmental problems. Good management practices are needed to achieve high productivity and healthy environmental conditions simultaneously. This requires close monitoring of environmental conditions and productivity, especially at lower trophic levels. Although environmental conditions, especially water quality, are widely monitored, primary production (PP) and secondary production (SP) are rarely monitored even though these basic parameters govern the structure and functioning of marine ecosystems. In this study, we developed methods to estimate PP and the SP from commonly collected water quality indicators and without the use of any information on phytoplankton or zooplankton except chlorophyll a (Chl.a) concentration. For this purpose, we developed the method to determine the half-saturation constants as a function of nutrient concentration, and the method to determine the SP from Chl.a concentration. Then, we used our newly developed methods to estimate the temporal distribution of PP and SP in the Seto Inland Sea from the 1980s to the present. To do so, we monitored water quality at three sites with different levels of eutrophication. We also measured PP by using 13C-labeled sodium bicarbonate (NaH13CO3) incubation method, and calculated copepod SP based on the species and body size of copepods collected in a survey. To estimate PP from water quality data, we modeled PP using a half-saturation constant for nutrient uptake that was allowed to vary as a function of nutrient concentration. Seasonal differences in PP could be successfully estimated by using this method. The estimation error for PP was within −16% to 23%. We found that SP calculated from copepod species and body size data was significantly correlated with chlorophyll a concentration. Therefore, we proposed a model to estimate SP from chlorophyll a concentration. Then, we used our newly developed models to estimate the temporal distribution of both PP and SP in the Seto Inland Sea from seasonal water quality data. According to our estimates, annual mean PP and SP peaked in the late 1990s and have trended downward since around 2000 or 2005. No significant inter-decadal changes in autumn or winter PP were observed during our study period; by contrast, spring and summer PP decreased significantly from the 1990s to the present. Estimated annual mean PP and SP over the most recent 5-year period were 73% and 88%, respectively, of equivalent values estimated for the 1990s.

In Situ Estimates of Net Primary Production in the Western North Atlantic With Argo Profiling Floats.

The 14C incubation method for net primary production (NPP) has limited spatial/temporal resolution, while satellite approaches cannot provide direct information at depth. With chlorophyll‐a and backscatter measurements from BGC‐Argo floats, we quantified year‐round NPP in the western North Atlantic Ocean using both the Carbon‐based Productivity Model (CbPM) and Photoacclimation Productivity Model (PPM). Comparison with NPP profiles from 14C incubation measurements showed advantages and limitations of both models. CbPM reproduced the magnitude of NPP in most cases. However, in the summer the CbPM‐based NPP had a large peak in the subsurface, which was an artifact from the subsurface chlorophyll maximum caused by photoacclimation. PPM avoided the artifacts from photoacclimation, but the magnitude of PPM‐derived NPP was smaller than the 14C result. Different NPP distribution patterns along a North‐South transect in the Western North Atlantic Ocean were observed, including higher winter NPP/lower summer NPP in the south, timing differences in NPP seasonal phenology, and different NPP depth distribution patterns in the summer months. Using a 6‐months record of concurrent oxygen and bio‐optical measurements from two Argo floats, we also demonstrated the ability of Argo floats to obtain estimates of the net community production to NPP ratio, ranging from 0.3 in July to −1.0 in December 2016. Our results highlight the utility of float bio‐optical profiles and indicate that environmental conditions (e.g., light availability, nutrient supply) are major factors controlling the seasonality and spatial (horizontal and vertical) distributions of NPP in the western North Atlantic Ocean.

Artificial propagation and larval breeding tests of the african sharptooth catfish, Clarias gariepinus conducted in captive conditions

The African catfish Clarias gariepinus is the second most cultured species in many African countries, including Senegal. Despite its economic importance in local aquaculture, artificial propagation of the species that would allow obtaining quality seed throughout the year has not yet been mastered adequately. This study aimed at producing C. gariepinus quality and mass supply seed to enhance local production. Three different types of incubation (Macdonald's bottle, water lettuce and wire mesh frame) were tested to identify the most efficient and accessible method for the producers at low cost and with a high hatching rate. The hatching rates obtained for the three different incubation methods were 26.9%, 10.0% and 6.3% for the Macdonald bottle (MB), wire frame (WF) and water lettuce (WL), respectively, while the corresponding survival rates were 91.6%, 70.4% and 77.8%.77.8%. The high hatching rate obtained with the MB could be due to the influence of permanent water circulation, which is absent in the other two hatching systems. The survival rates decreased 15 days after hatching and were 3.24%, 2.04% and 7.16%, respectively. The survival rates obtained three days after hatching were satisfactory, but the significant decrease at 15 days after hatching especially during the post- weaning phase seems to be due to the food quality, and unrelated to the hatching system. The daily individual growth, weight gain and body size of the larvae was significantly higher for larvae fed with natural food compared those that received artificial food. These results indicate that larvae preferred and fed the natural food which was permanently available in the rearing tanks. The artificial food deposited on the bottom of the tank changes appearance over time and may not be appreciated and efficiently consumed by the larvae, which may negatively impact their growth performance.

Comparison of three sperm selection methods for ICSI-DGC, Cumulus column, and incubation with supernatant product of adipose tissue-derived adult stem cells: An experimental study.

BackgroundThe examination of sperm parameters and sperm DNA integrity are necessary for male fertility expression. These parameters can be affected by method of sperm separation.ObjectiveTo measure the damage of each sperm separation method on the sperm parameters and sperm DNA integrity.Materials and MethodsIn this experimental study, semen samples of 20 infertile men with asthenoteratozoospermia (Infertility Research Center, Qom, Iran, 2017) were processed in three ways: density gradient centrifugation (DGC), cumulus column, and incubation with supernatant products of adipose tissue-derived adult stem cells (SPAS). The results of sperm parameters and DNA fragmentation before and after the process were statistically analyzed.ResultsThe number of separated sperms by normal morphologies during the SPAS and the cumulus column was significantly more than the corresponding population in the DGC group. In addition, although all three methods have the same ability to increase total sperm motility and the number of recovered sperms, in the field of forwarding movement and DNA fragmentation, the SPAS method performed more efficiently (p = 0.021).ConclusionSperm parameters and DNA fragmentation in the SPAS group were better than those in the DGC and cumulus column groups. Furthermore, it has been shown that the sperm capacity was increased with the SPAS method. However, the rearrangement of sperm chromatin by reducing the disulfide bridges and providing the possibility of re-histone over capacity causes a significant reduction in DNA fragmentation.

Husbandry and Captive Propagation of Eastern Collared Lizards (Crotaphytus collaris)

The eastern collared lizard (Crotaphytus collaris) is a colorful, common lizard found in the pet trade in the United States and Europe. As the popularity for the captive-bred collared lizard increases, reptile veterinarians should become more familiar with this native lizard species to best assist owners in providing appropriate husbandry, diet, and reproduction information. Recommended enclosures, lighting, temperature, shelter, and diet based on age are reviewed. We have also included comments regarding reproduction of collared lizards, including brumation, breeding and nesting, and recommended incubation methods. The few diseases currently reported for this genus, and a client care sheet, are also provided.

Mineralization of C and N in Whey-Treated Soils and Absorption of N by Plants

Acid whey deriving from milk processing is a nutrient-rich organic residue, with high potential for use in agriculture. However, in order to establish adequate application doses, it is paramount to understand its mineralization dynamics in the soil. The aims of this study were to evaluate C and N mineralization in two soils fertilized with acid whey and compare the efficiency of distinct incubation methods with and without leaching techniques in predicting the potentially mineralizable N of this residue. Samples of two soils (Haplustox and Haplustult) were submitted to five acid whey doses (0, 80, 160, 240, and 320 kg N ha−1), and the mineralization of C and N was determined. A pot experiment was conducted with the same soils and doses, in order to determine growth and N absorption by maize. C mineralization increased in both speed and quantity with the application of acid whey. Similarly, the potentially mineralizable N was increased, displaying mean values of 79.7 and 68.2 mg N kg−1 in the methods with and without leaching, respectively. N mineralization had strong correlation with absorbed N (r = 0.97 and 0.96, in the methods with and without leaching, respectively). The correlation between net mineralized N and absorbed N was higher without leaching (r = 0.98), in comparison to the leaching method (r = 0.89). Acid whey causes net N immobilization at the beginning of incubation, resulting in low N availability during this period, and its mineralization was better evaluated by the method without leaching.

Blood biocompatibility enhancement of biomaterials by heparin immobilization: a review.

Blood contacting materials are concerned with biocompatibility including thrombus formation, decrease blood coagulation time, hematology, activation of complement system, platelet aggression. Interestingly, recent research suggests that biocompatibility is increasing by incorporating various materials including heparin using different methods. Basic of heparin including uses and complications was mentioned, in which burst release of heparin is major issue. To minimize the problem of biocompatibility and unpredictable heparin release, present review article potentially reviews the reported work and investigates the various immobilization methods of heparin onto biomaterials, such as polymers, metals, and alloys. Detailed explanation of different immobilization methods through different intermediates, activation, incubation method, plasma treatment, irradiations and other methods are also discussed, in which immobilization through intermediates is the most exploitable method. In addition to biocompatibility, other required properties of biomaterials like mechanical and corrosion resistance properties that increase by attachment of heparin are reviewed and discussed in this article.