Long-term effects of cage-cleaning frequency and bedding type on laboratory rat health, welfare, and handleability: a cross-laboratory study

Cage-cleaning is necessary for a hygienic environment, but since rats communicate using scent, they might suffer if their cages are cleaned too frequently. Male rats (Sprague-Dawley and Wistar) were kept for five months across four animal units. Their cages were cleaned twice-weekly, weekly, or every two weeks, and contained either aspen woodchips or absorbent paper bedding. Aggression, injuries and general health, weight gain, chromodacryorrhoea (a stress-related Harderian gland secretion), handleability, and lung pathology were monitored, as was in-cage ammonia. Cleaning frequency had no clear impact on rat welfare, although frequent cleaning decreased ammonia concentrations and handleability, and non-aggressive skirmishing was highest in weekly cleaned rats. Surprisingly, bedding type did not affect ammonia, but all ammonia readings were unexpectedly low. However, rats kept on aspen had greater sneezing rates and lung pathology than those on paper bedding, but also had higher body weights. The results raise concerns about aspen bedding, which is relatively inert compared with other wood beddings, but nevertheless more harmful than paper. Animal unit significantly affected eight of the 11 variables tested, having interactive effects on five of them. The study also demonstrates the interactive effects of different animal units, casting doubt on the feasibility of standardization. We explored multiple variables of interest, so all findings require confirmation through further work. Nevertheless, cage-cleaning rates seem to affect socially housed male rats little, while bedding type has important effects on rat health.     

Effects of cage beddings on microsomal oxidative enzymes in rat liver

The purpose of the present studies was to evaluate the effects of some commercially available cage beddings on rat liver microsomal cytochrome P-450-dependent drug-metabolizing enzyme, ethylmorphine N-demethylase, and the carcinogen-metabolizing enzyme, benzo(a)pyrene hydroxylase. Sprague-Dawley rats were housed in cages containing cedar chip, corncob or heat-treated pinewood bedding for 3 weeks. Control rats were housed in cages on wire bottom floors containing no bedding material. Rats housed in cages containing cedar chip showed 18, 46 and 49% increases in liver cytochrome P-450 content, ethylmorphine N-demethylase and benzo(a)pyrene hydroxylase activities, respectively. The liver enzyme activities of rats housed in cages containing corncob bedding were similar to those obtained with control rats. In contrast, the pinewood-bedded rats showed a 21% decrease in ethylmorphine N-demethylase activity without affecting cytochrome P-450 content and benzo(a)pyrene hydroxylase activity. Hexobarbital-induced sleep times of the variously bedded rats were similar to those of control animals. These data suggest that the commercial bedding materials differ in their abilities to affect liver microsomal enzymes. Thus, interlaboratory variability in basal enzyme activities reported in the literature may be partly due to bedding materials used in the animal's cages.     

The bedding of laboratory animals as a source of airborne contaminants

In work environments with laboratory animals, the bedding of animals binds the excreta as well as other compounds originating from the animals and their environment. These may be generated into the ambient air when the personnel handle bedding in different procedures. This study compares the dustiness of different types of six clean and four soiled beddings from rat or mouse cages. The dust generation of clean bedding varied from <1 to 25 mg/m(3). When used in the cages of rats or mice for 4 days, the dust concentration of the beddings decreased, increased or stayed the same, depending on the type of bedding and animal species. A decrease in dustiness was, however, more common. The levels in the soiled beddings varied from <1 to 8.6 mg/m(3). In the case of the aspen chip bedding, the contents of bedding used in mouse, rat or rabbit cages were analysed for mesophilic bacteria and fungi, mycobacteria and endotoxins. All of these contaminants were variably found in the bedding samples, the maximal concentrations for bacteria were >6 500 000 colony-forming units (cfu)/g, for fungi 212 000 cfu/g, and for endotoxins 6500 ng/g (81 000 EU/g). The results showed that the bedding of laboratory animals may contain biologically effective compounds, and that these may be distributed into the ambient air depending on the characteristics of the bedding material. The dustiness of different bedding types is an important factor affecting the amount and quality of the occupational exposure of the personnel to airborne contaminants.     

Effect of experimental bedding treatments on the density of immature Musca domestica and Stomoxys calcitrans (Diptera: Muscidae) in outdoor calf hutches

Experimental bedding materials and a novel delivery method of cyromazine (Larvadex) were evaluated as replicated treatments in outdoor calf hutches for effect on the density of immature Musca domestica L. and Stomoxys calcitrans (L.). In 6-wk trials, overall density of Musca domestica L. and Stomoxys calcitrans (L.) in straw bedding averaged 36.2 and 52.6 maggots/liter, respectively, compared with respective average densities of 9.0 and 16.2 for wood chips and 10.4 and 20.0 for wood chips over a hydraulic fabric filter system. These values represent average reductions of 80 and 69% and 77 and 68%, respectively. Densities of M. domestica and S. calcitrans in ground corncob over the filter system averaged 13.3 and 4.1 maggots/liter in 3.2-mm size corncob particles, and 1.7 and 1.2 for 6.4-mm size corncob particles (average reductions of 62 and 87% and 90 and 96%, respectively) relative to straw bedding. Densities of these two species in sand bedding averaged 2.8 and 0.4 maggots/liter (average reductions of 93 and 99%), but sand became compacted and soiled with calf urine and feces after several weeks. Cyromazine excreted in calf urine limited density of both species in straw to an average of 11.0 and 15.6 maggots/liter, respectively (a 58 and 79% suppression relative to untreated straw). These data confirm that straw bedding promotes muscoid maggot growth and illustrate that alternative calf hutch bedding systems and urine delivery of cyromazine may improve muscoid fly management on dairy farms by limiting the development of muscoid maggots.     

Absorption of ethanol by steam-exploded corn stalk

The aim of this work is to study the feasibility of using a low-cost biomass absorbent steam-exploded corn stalk (SECS) to absorb ethanol in its production by fermentation. Measurement of many the physical properties of SECS showed its specific surface area was about 214 m(2)/g and it had a good structure for absorption. Some influencing parameters of using SECS to absorb ethanol in water were studied. Second-order and parabolic diffusion equations excellently described the kinetics of absorption for ethanol. Its absorption isotherm was well described by an improved BET equation, indicating that it was a process of polymolecular layer absorption and had phenomena similar to capillary coacervation. Mesh size did not significantly affect absorptivity, but absorbency decreased with temperature. Absorptivity of SECS for ethanol was compared to that of other absorbents: at 30 degrees C and 5% initial ethanol, the absorptivity of SECS for ethanol at 5h was 92 mg/g. When inactivated through use, SECS can continue to be used as an substrate to produce more ethanol, thus avoiding pollution through discarding.     

Evaluation of countermeasures for reduction of mouse airborne allergens

Three kinds of countermeasures for reduction of mouse airborne allergens were evaluated with use of an air sampler and immunochemical methods. Mouse cages and the sampler were placed inside a flexible-film isolator, and concentrations of mouse major allergens in the air were measured. The levels of the airborne allergens, prealbumin and albumin, generated by 10 males, were 3,050 and 492 pg/m3, respectively. Those by 10 females were lower, 317 and 270 pg/m3, respectively. When mouse cages were covered with a filter cap, the airborne allergens inside the isolator were decreased by 90%. When corncob was used as bedding in place of wood shavings, the airborne allergens were decreased by 57 and 77%, respectively. Therefore, for reduction of mouse airborne allergens, we recommend using female mice, covering the cages with filter caps, and using corncob bedding.     

阴离子型LDL吸附材料概述

动脉粥样硬化是导致心血管疾病发生的最重要因素。血液中低密度脂蛋白(LDL)浓度过高是引起动脉粥样硬化的主要原因。体外去除法是目前降低LDL浓度最有效的方法之一,吸附材料是影响LDL体外去除法降低LDL浓度效果的关键。阴离子型吸附材料是一种常用的吸附材料,因选材广泛、吸附效果佳备受关注,由发挥吸附功能的阴离子化合物配基和承载配基的载体基材组成,通过阴离子所带的负电荷与带正电的LDL产生特异性吸附。根据配基分子大小,阴离子型吸附材料主要分为大分子阴离子型和小分子阴离子型吸附材料,本文总结了国内外的阴离子型吸附材料主要研究现状及发展趋势。     

Effect of cage bedding on temperature regulation and metabolism of group-housed female mice

Mice are generally housed in groups in cages lined with an absorbent bedding material at ambient temperature (Ta) of 20 to 24 degrees C, which is comfortable for humans, but cool for mice. Little is known about the effects of bedding on thermoregulation of group-housed mice. To determine whether bedding material affects thermoregulatory stability, core temperature (Tc) and motor activity (MA) were monitored by use of radiotelemetry in female CD-1 mice housed in groups of four in a standard plastic cage at Ta of 23.5 degrees C. Ten groups were tested using three types of bedding material: a deep layer of heat-treated wood shavings (DWS) that allowed mice to burrow, a thin layer of wood shavings (TWS) just covering the bottom of the cage floor, or a layer of beta chips (BC). Mice could not burrow in the TWS or BC. The Tc and MA were affected by bedding type and time of day. Mice housed with DWS maintained a significantly higher Tc (deltaTc = 1.0 degrees C) during the day, compared with that in mice housed with TWS and BC. During the night, Tc and MA were high in all groups and there was no effect of bedding type on Tc or MA. Effect of bedding on metabolic rate (MR) was estimated by measuring oxygen consumption for six hours in groups of four mice at Ta of 23.5 degrees C. The Tc was significantly reduced in mice housed on the TWS and BC, but MR was unaffected by bedding type. There was a trend for higher MR in mice on BC. Compared with use of other bedding materials, housing mice on DWS and comparable materials provides an environment to burrow, thus reducing heat loss. The effects of bedding material on temperature regulation may affect rodent health and well being. Moreover, bedding will affect variability in toxicologic and pharmacologic studies whenever an endpoint is dependent on body temperature.     

The significance of differences in the mechanisms of photosynthetic acclimation to light, nitrogen and CO2 for return on investment in leaves

1. We report changes in photosynthetic capacity of leaves developed in varying photon flux density (PFD), nitrogen supply and CO₂ concentration. We determined the relative effect of these environmental factors on photosynthetic capacity per unit leaf volume as well as the volume of tissue per unit leaf area. We calculated resource-use efficiencies from the photosynthetic capacities and measurements of leaf dry mass, carbohydrates and nitrogen content.
2. There were clear differences between the mechanisms of photosynthetic acclimation to PFD, nitrogen supply and CO₂. PFD primarily affected volume of tissue per unit area whereas nitrogen supply primarily affected photosynthetic capacity per unit volume. CO₂ concentration affected both of these parameters and interacted strongly with the PFD and nitrogen treatments.
3. Photosynthetic capacity per unit carbon invested in leaves increased in the low PFD, high nitrogen and low CO₂ treatments. Photosynthetic capacity per unit nitrogen was significantly affected only by nitrogen supply.
4. The responses to low PFD and low nitrogen appear to function to increase the efficiency of utilization of the limiting resource. However, the responses to elevated CO₂ in the high PFD and high nitrogen treatments suggest that high CO₂ can result in a situation where growth is not limited by either carbon or nitrogen supply. Limitation of growth at elevated CO₂ appears to result from internal plant factors that limit utilization of carbohydrates at sinks and/or transport of carbohydrates to sinks.     

Use of available crop by-products as alternative bedding materials to wheat straw for rearing broilers

The provision of suitable bedding for rearing broilers is an ongoing challenge for poultry producers. The aim of the present study was to evaluate the effect of using different types of crop residues as alternative bedding materials to wheat straw on growth performance, carcass traits, health, and welfare of broilers; moreover, the effects on the bedding and air quality were analyzed. We randomly assigned 180 1-day-old chickens (Cobb 500) across six bedding groups (three replicates of 10 birds each). Chickens in all groups were reared under similar housing conditions. Chickens of one group were reared on a wheat straw bedding (control), whereas those of the remaining five groups were reared on clover straw, cornstalk chips (CS), sugarcane top chips, chopped palm spines (CPS), and corn ear husks. Cornstalk chip and chopped palm spines were associated with the lowest (P < 0.05) ammonia concentrations compared with the remaining beddings. The mean values of bedding moisture percentage (P < 0.05), caking score, and ammonia concentrations (P < 0.05) were decreased with 6-week-old broilers reared on cornstalk chips. At 6 weeks, wheat straw, clover straw, sugarcane top chips and corn ear husks resulted in higher (P < 0.0001) BW and average daily gain compared with CS and CPS. Broilers reared on clover straw bedding showed the best FCR during 4–6 weeks of age (P < 0.01), and those kept on clover straw and sugarcane top chips exhibited the best (P < 0.01) feed conversion ratio (1.61 and 1.64, respectively) during 0–6 weeks of age. The carcass dressing yield, airborne dust concentration, bedding pH, and bacterial counts did not differ among the different bedding materials. The proportions of the drumsticks, gizzards, and abdominal fat of 6-week-old broilers were affected by bedding type (P < 0.05); but the differences among the bedding treatments were small. The mean incidence of footpad dermatitis and breast blisters was significantly less using clover straw. Broilers reared on chopped palm spine had the highest (P < 0.05) heterophil/lymphocyte ratio when compared with their counterparts. Conclusively, using clover straw or cornstalks as an alternative bedding material may be a beneficial strategy and should be highly recommended for rearing broilers.     

Dust and ammonia in horse stalls with different ventilation rates and bedding

Airborne particles and ammonia were monitored in horse stalls managed under four conditions. Two ventilation rates, high (27 air changes per h) and low (5 air changes per h) and two bedding types, paper and straw, were employed. At both ventilation rates, the number of airborne particles generated while the stalls were mucked out was higher with straw than with paper. Particles were more efficiently cleared at the higher ventilation rate in both the straw and paper stalls. Ammonia measurements reflected an accumulation over time. In the stalls with low ventilation, ammonia levels were significantly higher than in those stalls with high ventilation regardless of bedding type. Management decisions and their relationships to respiratory disorders are discussed.     

Dust and ammonia in horse stalls with different ventilation rates and bedding

Airborne particles and ammonia were monitored in horse stalls managed under four conditions. Two ventilation rates, high (27 air changes per h) and low (5 air changes per h) and two bedding types, paper and straw, were employed. At both ventilation rates, the number of airborne particles generated while the stalls were mucked out was higher with straw than with paper. Particles were more efficiently cleared at the higher ventilation rate in both the straw and paper stalls. Ammonia measurements reflected an accumulation over time. In the stalls with low ventilation, ammonia levels were significantly higher than in those stalls with high ventilation regardless of bedding type. Management decisions and their relationships to respiratory disorders are discussed.     

Partitioning of metabolizable energy intake in sucking altricial and precocial rodent pups

We partitioned the metabolizable energy intake (MEI) into energy for maintenance (ME_m) and for growth (ME_g) in sucking precocial and altricial rodent pups. Each of the two components includes energy loss due to the heat increment of feeding. ME_m of precocial pups expressed as average daily energy costs or as a proportion of MEI was greater than in altricial pups of similar size and, therefore, less energy was available for growth. Consequently, the overall energy cost (via total MEI) per unit postnatal growth of precocial pups was greater than for altricial pups of similar size. We used the proportion of calculated ME_m to that predicted by body mass as an index of precociality in rodent pups. The proportion of ME_g to MEI in precocial pups was lower than in altricial pups and was inversely related to the index of precociality.     

Effect of animal bedding on rat liver endosome acidification

Animal beddings, such as pine products, and environmental factors are known to induce liver drug-metabolizing cytochrome P450 enzymes. We observed that a change to pine-based rat bedding altered baseline and cAMP-stimulated rates of acidification in rat liver endosomes, apparently by decreasing ATP-dependent proton transport in the presence and absence of chloride. Although cAMP altered phosphorylation of protein kinase B and extracellular signal-regulated kinases 1 and 2 (ERK 1,2) and p38 mitogen-activated protein kinases, changes in housing conditions did not affect baseline or cAMP-stimulated values of these or other selected signaling molecules. We conclude that compounds in rat bedding may alter not only drug metabolism, but also aspects of endocytosis.     

Biodegradable superabsorbent hydrogels derived from cellulose by esterification crosslinking with 1,2,3,4-butanetetracarboxylic dianhydride

Superabsorbent hydrogels were prepared from native celluloses dissolved in lithium chloride and N-methyl-2-pyrrolidinone (LiCl/NMP) by esterification crosslinking with 1,2,3,4-butanetetracarboxylic dianhydride (BTCA). Subsequent conversion of the unreacted carboxyl groups to sodium carboxylates by the addition of aqueous NaOH was performed to enhance the water affinity of the gels. The absorbency of the products was strongly dependent on the amount of BTCA that was esterified to cellulose, and the highest absorbency was observed for the hydrogel composed of approximately 0.25 molecules of BTCA per anhydroglucose unit (AGU) of cellulose. Furthermore, it was confirmed that the absorbency was enhanced as the average degree of polymerization (DP) of the starting cellulose increased. The use of cotton cellulose with a high DP of about 2400 produced a hydrogel with an absorbency of 720 times its dry weight, which exceeded the absorbency of commercial crosslinked sodium polyacrylate superabsorbent hydrogel (SPA). The hydrogels exhibited good biodegradability, with a maximum degradation of 95% within 7 days using cellulase.     

Evidence of thermostable amylolytic activity from Rhizopus microsporus var. rhizopodiformis using wheat bran and corncob as alternative carbon source

Rhizopus microsporus var. rhizopodiformis produced high levels of alpha-amylase and glucoamylase under solid state fermentation, with several agricultural residues, such as wheat bran, cassava flour, sugar cane bagasse, rice straw, corncob and crushed corncob as carbon sources. These materials were humidified with distilled water, tap water, or saline solutions--Segato Rizzatti (SR), Khanna or Vogel. The best substrate for amylase production was wheat bran with SR saline solution (1:2 v/v). Amylolytic activity was still improved (14.3%) with a mixture of wheat bran, corncob, starch and SR saline solution (1:1:0.3:4.6 w/w/w/v). The optimized culture conditions were initial pH 5, at 45 degrees C during 6 days and relative humidity around 76%. The crude extract exhibited temperature and pH optima around 65 degrees C and 4-5, respectively. Amylase activity was fully stable for 1 h at temperatures up to 75 degrees C, and at pH values between 2.5 and 7.5.     

Longitudinal measurements of blood volume and essential body mass in human subjects.

Repeated determinations of blood volume and body density were made on 34 Chinese subjects (28 men and 6 women) in Taiwan over a period of 12 yr, as the mean age increased from 31 to 43 yr. Essential body mass calculated from body density and body weight showed no significant change over the 12-yr period. Changes in body weight (mean gain equals 6.0 kg) were attributable to alterations in adipose tissue weight, the density of which was found to be 0.948 g/cm3. In two-thirds of the subjects the second blood volume increased by more than 5% over the first determination, and the mean blood volume for all subjects increased by 7.5% (P less than 0.01). Correlation of the blood volume data with the findings on essential body mass and adipose tissue mass suggests that blood content per unit tissue mass increased in the second determination. This interpretation is supported by the increase in nutrient availability in Taiwan over the 12-yr period, and it may explain the lack of blood volume increase in an earlier longitudinal study on American subjects with stable nutrient availability.     

Relationships between photosynthesis, nitrogen and leaf structure in 14 grass species and their dependence on the basis of expression

The relationships between leaf structure, nitrogen concentration and CO₂ assimilation rate ( A ) were studied for 14 grass species grown in the laboratory under non-limiting nutrient conditions. Structural features included leaf thickness and density, and the proportion of leaf volume occupied by different types of tissue (mesophyll, epidermis, vessels and sclerenchyma). Relationships were assessed for data expressed per unit leaf area and fresh mass. The latter was found to be closely related to leaf volume, which allowed us to use A per unit leaf fresh mass ( A _fm) as a surrogate of A per unit leaf volume. Assimilation rate per unit leaf area ( A _a) was positively correlated with leaf thickness and with the amount of mesophyll per unit leaf area; the relationship with leaf nitrogen content per unit area was only marginally significant. A _fm was negatively correlated with leaf thickness and positively with fresh mass-based leaf organic nitrogen concentration. A multiple regression involving these two variables explained 81% of the variance in A _fm. The value of A _fm was also significantly related to the proportion of mesophyll in the leaf volume, but surprisingly the correlation was negative. This was because thin leaves with high A _fm and nitrogen concentration had proportionally more mechanically supportive tissues than thick ones; as a consequence, they also had a lower proportion of mesophyll. These data suggest that, in addition to leaf nitrogen, leaf thickness has a strong impact on CO₂ assimilation rate for the grass species studied.     

Synthesis and characterization of a novel super-absorbent based on wheat straw

In order to develop an eco-friendly polymer, a novel super-absorbent polymer was prepared by graft copolymerization of acrylic acid (AA), acrylic amide (AM) and dimethyl diallyl ammonium chloride (DMDAAC) onto the pretreatment wheat straw (PTWS). The molecular structure of the super-absorbent was confirmed by FTIR. The factors that can influence absorbencies of the super-absorbent resin (SAR) were investigated, such as weight ratio between the monomers, the ratio of PTWS to monomers, the amount of initiator and cross-linker, temperature reaction time and neutralization degree of AA. The SAR has the water absorbency of 133.76 g/g in distilled water and 33.83 g/g in 0.9 wt.% NaCl solution.     

Impact of proliferative activity and tumorigenic conversion on mitochondrial function of fibroblasts in 2D and 3D culture

The purpose of the present study was to examine mitochondrial function in differently transformed cells relative to their tumorigenic state and proliferative activity in vitro. An established two-step carcinogenesis model consisting of immortal and tumorigenic rat embryo fibroblasts that can be cultured as monolayers and multicellular spheroids was investigated. Flow cytometric measurements were carried out using the two mitochondrial-specific fluorochromes rhodamine 123 (Rh123) and 10-N-nonyl acridine orange (NAO), in combination with the DNA dye Hoechst 33342 for simultaneous cell cycle analysis. Since the accumulation of Rh123 depends on mitochondrial membrane potential, Rh123 fluorescence intensity gives an estimate of mitochondrial activity per cell, as determined by both overall mitochondrial function and mass. In contrast, NAO uptake reflects mitochondrial mass only, as it binds to cardiolipin in the inner mitochondrial membrane independently of membrane potential. Aliquots of cell suspensions derived from exponential monolayer, confluent monolayer, and a range of sizes of multicellular spheroids were stained with either Rh123 or NAO and Hoechst 33342, then mitochondrial mass and activity per unit cell volume and cellular DNA content were measured by flow cytometry. Differences in the average mitochondrial activity per cell in different cell lines and culture conditions were primarily due to alterations in cell volume. Importantly, tumorigenic conversion by ras-transfection did not consistently change mitochondrial activity per unit cell volume. The mitochondrial mass per unit cell volume increased for all cells when cellular quiescence was induced, either in monolayers or spheroids. However, mitochondrial function (activity/mass) decreased when cells became quiescent, resulting in a positive correlation between mitochondrial function and S-phase fraction, independent of transformation status or culture condition. We conclude that mitochondrial function reflects proliferative activity rather than tumorigenic conversion.