Cloning and characterization of an antifungal class III chitinase from suspension-cultured bamboo ( Bambusa oldhamii ) cells

A class III chitinase cDNA (BoChi3-1) was cloned using a cDNA library from suspension-cultured bamboo ( Bambusa oldhamii ) cells and then transformed into yeast ( Pichia pastoris X-33) for expression. Two recombinant chitinases with molecular masses of 28.3 and 35.7 kDa, respectively, were purified from the yeast's culture broth to electrophoretic homogeneity using sequential ammonium sulfate fractionation, Phenyl-Sepharose hydrophobic interaction chromatography, and Con A-Sepharose chromatography steps. N-Terminal sequencing and immunoblotting revealed that both recombinant chitinases were encoded by BoChi3-1, whereas SDS-PAGE and glycoprotein staining showed that the 35.7 kDa isoform (35.7 kDa BoCHI3-1) was glycosylated and the 28.3 kDa isoform (28.3 kDa BoCHI3-1) was not. For hydrolysis of ethylene glycol chitin (EGC), the optimal pH values were 3 and 4 for 35.7 and 28.3 kDa BoCHI3-1, respectively; the optimal temperatures were 80 and 70 degrees C, and the K(m) values were 1.35 and 0.65 mg/mL. The purified 35.7 kDa BoCHI3-1 hydrolyzed EGC more efficiently than the 28.3 kDa isoform, as compared with their specific activity and activation energy. Both recombinant BoCHI3-1 isoforms showed antifungal activity against Scolecobasidium longiphorum and displayed remarkable thermal (up to 70 degrees C) and storage (up to a year at 4 degrees C) stabilities.     

Elimination behavior patterns of domestic cats (Felis catus) with and without elimination behavior problems

OBJECTIVE: To investigate the relationship of litter box location as it relates to cats' use of space in the house, elimination problems, and certain behaviors associated with elimination. SAMPLE POPULATION: 40 cats in single-cat house-holds with or without elimination behavior problems (20 cats/group). PROCEDURES: Camcorders were used to record the cats' behaviors at the litter box and other areas in which they eliminated during a 72-hour period. Use of space in the house was recorded by direct observation during 400 minutes of the 72-hour period. Elimination behaviors and other cat- and litter box-associated variables were compared between groups; litter box location with respect to inappropriate elimination was assessed. RESULTS: Litter box location did not differ between cats with and without elimination behavior problems. An inverse correlation was found between time spent sniffing and the distance of the litter box from the central core area. Cats with elimination problems spent significantly less time digging at the litter box than cats without elimination problems. There was no significant difference in the time spent pawing in litter box, sniffing, or covering excreta after elimination between the 2 groups of cats. CONCLUSIONS AND CLINICAL RELEVANCE: Times spent digging in litter boxes by cats with and without elimination problems have been determined, and data suggest that actual digging times could be used as a means to test for litter preference and litter aversion. This information may also be used to identify cats with litter aversion prior to the development of an elimination problem.     

Impacts of weaning age on dietary needs of whey permeate for pigs at 7 to 11 kg body weight

Background:Whey permeate is an effective lactose source for nursery pigs and the most benefits are obtained when pigs are at 7 to 11 kg BW.Altering weaning ages could cause different length of early-weaner phases until 7 kg BW and thus it would influence the dietary need of whey permeate during 7 to 11 kg BW of pigs.This study aimed to evaluate if weaning ages would affect the dietary needs of whey permeate for optimum growth performance of pigs at 7 to 11 kg BW.Methods:A total of 1,632 pigs were weaned at d 21(d 21.2±1.3)or d 25(d 24.6±1.1)after birth.All pigs had a common early-weaner feeds until they reached 7 kg BW.When pigs reached 7 kg BW within a weaning age group,they were allotted in a randomized complete block design(2×4 factorial).Two factors were weaning age groups(21 and 25 d of age)and varying whey permeate levels(7.50%,11.25%,15.00%,and 18.75%).Data were analyzed using the GLM and NLIN procedures of SAS for slope-ratio and broken-line analyses to determine the growth response to whey permeate and optimal daily whey permeate intake for the growth of the pigs weaned at different ages.Results:Pigs weaned at 21 d of age had a common diet for 11 d to reach 7 kg BW whereas pigs weaned at 25 d of age needed 2 d.The G:F of pigs weaned at 25 d of age responded to increased daily whey permeate intake greater(P<0.05)than pigs weaned at 21 d of age.Breakpoints were obtained(P<0.05)at 88 and 60 g/d daily whey permeate intake or 17.0%and 14.4%of whey permeate for G:F of pigs weaned at 21 and 25 d of age,respectively.Conclusion:Pigs weaned at an older age with a short early-weaner phase had a greater growth response to whey permeate intake compared with pigs weaned at a younger age with a long early-weaner phase.Altering weaning ages affected dietary needs of whey permeate for optimum growth performance of pigs from 7 to 11 kg BW.     

Combined effect of biochar and soil moisture on soil chemical properties and microbial community composition in microplastic-contaminated agricultural soil

Accumulation of microplastics (MPs) in agricultural environments has caused growing concern in recent years because of its detrimental impacts on soil quality, crop productivity and ecosystem function. This study was conducted to assess the impact of biochar on soil chemical and microbial properties in a MP-contaminated soil under two moisture regimes. Soil was contaminated with 1% (w/w) of low-density polyethylene MPs. Four types of standard biochar, that is, oil seed rape (OSR) biochar produced at 550°C (OSR 550) and 700°C (OSR 700) and soft wood pellet (SWP) biochar produced at 550°C (SWP 550) and 700°C (SWP 700), were applied at a rate of 5% (w/w). The control was maintained without MP addition. The samples were incubated in soil with two moisture regimes, that is, at 30% and 70% of the water holding capacity, and the soil chemical and microbiological properties were assessed after 100 days of incubation. OSR biochar application significantly increased soil pH (8.53–8.81) and electrical conductivity (0.51–0.58 dS/m) in both moisture regimes. The effect of biochar application on soil enzyme activity and microbial community composition did not show a clear trend. However, SWP 700 biochar improved soil enzyme activity compared with that of the control and improved bacterial diversity and evenness compared with those of other biochars, which was attributed to the high surface area available for microbial colonization. Low soil moisture content significantly reduced enzyme activity and bacterial richness even with biochar amendment, except for SWP 550 biochar. This study implies the suitability of biochar for improvement of soil quality in MP contaminated soil under both moisture regimes. However, further long-term studies are needed to get a clear understanding on the impact of different types of biochar on MP-contaminated soil.