Red galls: the different stories of two gall types on the same host

• Several studies have suggested reasons why galls have conspicuous colours, but none of the ideas have been confirmed. However, what if the vibrant colours of some galls are explained simply by the effect of light exposure? This may lead to anthocyanin accumulation, functioning as a defence mechanism against the effects of high light.
• We studied the globoid galls induced by Cecidomyiidae (Diptera) on Qualea parviflora (Vochysiaceae), relating anthocyanin accumulation and chlorophyll fluorescence parameters to light incidence in abaxial and adaxial galls. We also tested if the anthocyanin accumulation patterns apply to another Cecidomyiidae‐induced gall morphotype (intralaminar) within the same plant.
• Adaxial galls are exposed to higher incident light, with more anthocyanin accumulation and therefore red coloration. In galls from angled leaves, the greater the angle of the leaf, the higher the difference between anthocyanins on the sun and shade sides of galls. Photosynthetic pigment concentrations did not differ between abaxial and adaxial galls. However, we found higher (F_m′ ? F′)/F_m′ and F_v/F_m in the abaxial galls. Conversely, NPQ and R_fd were higher in adaxial galls. Finally, the pattern of anthocyanin accumulation was not found in the intralaminar gall.
• Anthocyanin accumulation in galls functions as a photoprotective strategy, maintaining tissue vitality in regions exposed to high light conditions. However, this mechanism may vary even among galls within the same host, indicating idiosyncrasy when it comes to coloration in galls. To date, this is the first study to demonstrate quantitatively why the galls of a specific species may be coloured: the variation in light regimes creates differential anthocyanin accumulation, influencing coloration.

     

Influence of environmental conditions on infection of Klebsiella pneumoniae by two different types of bacteriophages

The adsorption and efficiency of plating of bacteriophages FC3-1 and FC3-9 on Klebsiella pneumoniae C3 (serotype O1:K66) cells grown at different pHs and temperatures were quantitated. Bacteriophage FC3-1, with lipopolysaccharide as its bacterial receptor, showed a large decrease in efficiency of plating on bacteria grown at low pH or low temperature. Under the same conditions, no significant decrease in efficiency of plating was found for bacteriophage FC3-9, a phage requiring capsule and lipopolysaccharide for its adsorption and carrying capsule-depolymerizing activity. We demonstrate that K. pneumoniae C3 cells grown at low pH or low temperature have less lipopolysaccharide exposed on their surface. We conclude that this is why lipopolysaccharide-specific phage FC3-1 less efficiently infects bacterial cells grown under those conditions. We propose that bacteriophage FC3-9 efficiently infects bacterial cells grown at low pH or low temperature because its enzymatic activity on the capsule makes lipopolysaccharide available to this phage.     

Comparison of survival rates in four inbred mouse strains under different housing conditions: effects of environmental enrichment

Housing conditions can affect the well-being of laboratory animals and thereby affect the outcomes of experiments. The appropriate environment is essential for the expression of natural behavior in animals. Here, we compared survival rates in four inbred mouse strains maintained under three different environmental conditions. Three mouse strains (C57BL/6J, C3H/HeN, and DBA/2J) housed under environmental enrichment (EE) conditions showed improved survival; however, EE did not alter the survival rate of the fourth strain, BALB/c. None of the strains showed significant differences in body weights or plasma corticosterone levels in the three environmental conditions. For BALB/c mice, the rates of debility were higher in the EE group. Interestingly, for C57BL/6J and C3H/HeN mice, the incidence of animals with alopecia was significantly lower in the EE groups than in the control group. It is possible that the enriched environment provided greater opportunities for sheltering in a secure location in which to avoid interactions with other mice. The cloth mat flooring used for the EE group was bitten and chewed by the mice. Our findings suggest that depending on the mouse strains different responses to EE are caused with regard to health and survival rates. The results of this study provide basic data for further studies on EE.     

Assessment of welfare of finishing beef cattle kept on different types of floor after short- or long-term housing

This study aimed at evaluating short- and long-term effects of housing beef cattle on deep litter (DL) or concrete fully slatted floor (FS) on their welfare. Animal-based measures of the Welfare Quality® assessment protocol for cattle were used to assess health status and behaviour of bulls. The assessment was carried out in a large commercial farm on 15 batches of bulls (4 DL and 11 FS) 1 month after their receiving day (short-term) and on 12 batches (three DL and nine FS) the week before slaughter (long-term). Signs of better comfort on deep litter in terms of shorter lying down durations (5.1±0.5 v. 6.5±0.4 s; P<0.05) and lower risk of hairless patches (odds ratio=0.09; 95% confidence interval=0.01 to 0.68; P<0.05) were already observed after 1 month. Heavy bulls after a long-term housing on FS showed a higher prevalence of bursitis, hairless patches and lesions/swellings than animals on DL. Bulls on fully slatted floor were at higher risk of early culling (odds ratio=6.44; 95% confidence interval=1.57 to 26.37; P<0.01), mainly due to musculoskeletal system pathologies/lameness. Deep litter proved to be a valid alternative to slatted floor, making animals more confident to interact with powerful movements such as mounting at the end of the finishing period. A negative aspect of the deep litter was the poor cleanliness of the bulls. Compared with the fully slatted floor, there were higher odds ratios for dirty bulls at both, the short- (odds ratio=25.09; 95% confidence interval=8.96 to 70.22; P<0.001) and the long-term housing (odds ratio=276.13; 95% confidence interval=98.21 to 776.38; P<0.001). In order to improve health and welfare of beef cattle finished at a heavy weight, deep litter systems are a promising alternative to fully slatted floors. However, proper management of deep litter is necessary to maintain satisfactory cleanliness of the bulls.     

The welfare of growing pigs in five different production systems: assessment of feeding and housing

Ninety-one farms were visited over a 2-year period to assess the welfare of growing pigs in five different production systems found either in France or in Spain using the Welfare Quality® protocol. This study focused on animal-based measures as indicators of ‘good feeding’ and ‘good housing’. Multiple Generalized Linear Mixed Models were performed for each measure to evaluate the differences between production systems and to detect possible causal factors. Pigs in the conventional system presented the lowest prevalence of poor body condition, whereas extensive Mallorcan Black pigs and extensive Iberian pigs were associated with a decreased prevalence of bursitis and pig dirtiness. The straw-bedded system presented a lower prevalence of bursitis, but poorer hygiene and more susceptibility of poor body condition than the conventional system. The age of the animals had a significant effect on the appearance of bursitis in the three intensive systems studied. The type of floor was a significant causal factor of bursitis and pig dirtiness in the conventional system and among intensive Iberian pigs. The feeding system was another causal factor of pig dirtiness on more than 50% of the body in the conventional system, whereas pig dirtiness on less than 50% of the body was influenced by the age of the animals. The prevalence of huddling animals in the conventional system was associated with the highest stocking densities and the lowest environmental temperatures. The results indicate that there were important differences between production systems based on animal-based indicators of the good feeding and housing principles. The recording of the age of the animals, type of floor, feeding system, stocking density and environmental temperature can be useful to predict the appearance of a given welfare measure of ‘good housing’ on a farm.     

Impact of environmental enrichment in mice. 1: effect of housing conditions on body weight,organ weights and haematology in different strains

Currently, environmental enrichment is a very common means of improving animal well-being, especially for laboratory animals. Although environmental enrichment seems to be a possible way for improving the well-being of animals, the consideration of housing laboratory animals should not only focus solely on animal well-being, manpower and economics but also on the precision and accuracy of the experimental results. The purpose of the present study was to evaluate the effects of enriched cages (nest box, nesting material, climbing bar) on body weight, haematological data and final organ weights. BALB/c, C57BL/6 and A/J mice, originated from Harlan Winkelmann, were used for the experiments - 16 animals of each strain. Animals at 3 weeks of age were marked and separated randomly to enriched or non-enriched cages, in groups of four, half for each housing condition. Both cages were type III Makrolon cages, only the enriched cages contained a nest box, a wood bar for climbing and nesting material. Animals were kept in a clean animal room under specific pathogen free (SPF) conditions. Body weights were recorded every week. Blood samples were collected at 14 weeks of age (white blood cells (WBC), red blood cells (RBC), haemoglobin (HGB), and haematocrit (HCT) were analysed). At 15 weeks of age, the animals were euthanized by CO(2) in their home cages, and final body weight and organ weights (heart, liver, kidney, adrenal, spleen and uterus) were recorded immediately. Although nearly all the test variables were not affected by environmental enrichment in their mean values, the enriched group showed higher coefficients of variation in many variables, and strain differences of both housing conditions were not consistent. The influences of enrichment were shown to be strain- and test-dependent. Such effects may lead to an increase in the number of animals which is necessary or may change the experimental results, especially when a study, using enriched housing conditions, focuses on strain differences. Since the same enrichment design can result in different influences, a positive or a negative or no adverse effect, due to the strain and the variables studied, researchers need to collect more information before enrichment designs are introduced into experimental plans.     

Influence of vitamins A, D3 and E status on post-mortem meat quality in steers under winter housing or pasture finishing systems

We investigated the influence of Swedish recommended vitamins A, D3 and E supplementation levels on muscle tenderness and fatty acid (FA) composition under indoor or outdoor finishing programmes. Swedish Red breed steer calves were divided into vitamin supplemented (n = 12) and non-supplemented (n = 15) groups while on pasture prior to the finishing period. This trial began at the beginning of the winter housing period during which the steers were fed a 55 : 45 dry matter barley : grass silage diet indoors. The indoor finished group was comprised of vitamin supplemented (n = 6) and non-supplemented (n = 8) steers slaughtered after about 155 days on feed. Vitamin supplemented steers were provided with 100 g mineral supplement providing 400 000 IU vitamin A, 100 000 IU D3 and 3000 IU E daily as recommended for Swedish production practices. In spring, outdoor finished vitamin supplemented (n = 6) and non-supplemented (n = 7) steers grazed semi-natural grassland for an additional 120 days before slaughter. During pasture, vitamin supplemented steers had free-choice access to a mineral supplement containing vitamins A, D3 and E. The mineral supplement for the non-supplemented steers did not contain vitamins A, D3 and E and was provided at the same amount as the vitamin supplemented steers. Shear force values were similar between vitamin supplemented and non-supplemented steers after ageing 2, 7 and 14 days within indoor and outdoor finishing programmes. The shear force values had decreased by 14 days of ageing within all programmes. The μ- and m-calpain activity did not differ between vitamin supplemented and non-supplemented steers for either the indoor or outdoor finishing programmes. The calpastatin activity was higher for the indoor, vitamin supplemented steers. Indoor finished vitamin supplemented steers had a greater proportion of C18:1c-9 and total monounsaturated fatty acids, whereas the non-supplemented steers had a greater proportion of total saturated fatty acids. We concluded that the meat quality from steers not receiving vitamin supplementation was similar to that of steers receiving vitamins A, D3 and E supplementation at Swedish recommended levels under indoor and outdoor finishing programmes.     

The two types of 3-dehydroquinase have distinct structures but catalyze the same overall reaction.

The structures of enzymes catalyzing the reactions in central metabolic pathways are generally well conserved as are their catalytic mechanisms. The two types of 3-dehydroquinate dehydratase (DHQase) are therefore most unusual since they are unrelated at the sequence level and they utilize completely different mechanisms to catalyze the same overall reaction. The type I enzymes catalyze a cis-dehydration of 3-dehydroquinate via a covalent imine intermediate, while the type II enzymes catalyze a trans-dehydration via an enolate intermediate. Here we report the three-dimensional structures of a representative member of each type of biosynthetic DHQase. Both enzymes function as part of the shikimate pathway, which is essential in microorganisms and plants for the biosynthesis of aromatic compounds including folate, ubiquinone and the aromatic amino acids. An explanation for the presence of two different enzymes catalyzing the same reaction is presented. The absence of the shikimate pathway in animals makes it an attractive target for antimicrobial agents. The availability of these two structures opens the way for the design of highly specific enzyme inhibitors with potential importance as selective therapeutic agents.     

Effect of two types of environmental enrichment for singly housed mice on food intake and weight gain

For animals that cannot be housed socially for scientific reasons, such as rodents on food intake-energy expenditure studies, the provision of environmental enrichment may alleviate stress. However, the influence of environmental enrichment on food intake and energy expenditure is unresolved. The authors tested the effects of two environmental enrichment options on food intake and weight gain in singly housed mice. Their results may be helpful in developing rodent enrichment programs.     

Effects of environmental enrichment and regrouping on natural autoantibodies-binding danger and neural antigens in healthy pigs with different individual characteristics

Pigs living in commercial husbandry systems may experience both acute stress due to standard management procedures and chronic stress through limitations in their barren housing environment. This might influence their immune status, including antibody responses to neural and danger autoantigens. Levels of natural autoantibody (NAAb)-binding phosphorylcholine-conjugated bovine serum albumin (PC-BSA) and myelin basic protein (MBP) were measured over time in pigs that were kept in environmental enriched v. barren housing, and that underwent a regrouping test. In total, 480 pigs were housed in 80 pens in either barren or straw-enriched pens from 4 through 23 weeks of age. Blood samples were taken from pigs before (week 8), and 3 days after a 24 h regrouping test (week 9), and at 22 weeks of age. Phosphorylcholine-conjugated bovine serum albumin (PC-BSA) and MBP antibody titres in serum were measured using ELISA. Enriched-housed pigs had higher levels of IgM-binding MBP, and tended to have higher levels of IgG-binding MBP and IgA-binding PC-BSA than barren-housed pigs. Each NAAb measured in this study was affected by gender and litter. These results suggest that enriched housing conditions, as well as acute regrouping stress, have an influence on levels of serum NAAb-binding danger and neural antigens in pigs.     

Neuronal nicotinic acetylcholine receptors from Drosophila: two different types of alpha subunits coassemble within the same receptor complex

Although neuronal nicotinic acetylcholine receptors from insects have been reconstituted in vitro more than a decade ago, our knowledge about the subunit composition of native receptors as well as their functional properties still remains limited. Immunohistochemical evidence has suggested that two alpha subunits, alpha-like subunit (ALS) and Drosophila alpha2 subunit (Dalpha2), are colocalized in the synaptic neuropil of the Drosophila CNS and therefore may be subunits of the same receptor complex. To gain further understanding of the composition of these nicotinic receptors, we have examined the possibility that a receptor may imbed more than one alpha subunit using immunoprecipitations and electrophysiological investigations. Immunoprecipitation experiments of fly head extracts revealed that ALS-specific antibodies coprecipitate Dalpha2, and vice versa, and thereby suggest that these two alpha subunits must be contained within the same receptor complex, a result that is supported by investigations of reconstituted receptors in Xenopus oocytes. Discrimination between binary (ALS/beta2 or Dalpha2/beta2) and ternary (ALS/Dalpha2/beta2) receptor complexes was made on the basis of their dose-response curve to acetylcholine as well as their sensitivity to alpha-bungarotoxin or dihydro-beta-erythroidine. These data demonstrate that the presence of the two alpha subunits within a single receptor complex confers new receptor properties that cannot be predicted from knowledge of the binary receptor's properties.     

Comparison of lymphocyte immunophenotypes obtained simultaneously from two different data acquisition and analysis systems on the same flow cytometer.

Immunophenotyping of different lymphocyte populations was carried out in parallel on 113 consecutively received specimens of human peripheral blood using 2 different data acquisition and analysis systems (EPICS C and 4Cyte-Acmecyte) on the same flow cytometer (EPICS C). The phenotypes analyzed were CD3+, CD4+, CD8+ CD56+ CD16+ CD3-, TCR-gamma delta+ CD8-, and TCR-gamma delta+ CD8+. Both HIV- and HIV+ specimens were used for this study, including some with CD4 levels as low as 2% of all lymphocytes. Despite differences in gating procedures and shapes of bitmap (rectilinear vs. "amorphous"), the 2 methods agreed to within 2% positive cells in 97% of the cases. Although some statistically significant biases in the methods were observed, these were small and not biologically important. We conclude that both methods of data acquisition and analysis, as employed by experienced operators on the EPICS C flow cytometer, gave essentially equivalent results for lymphocyte sub-populations in peripheral blood preparations.     

Reflection versus fluorescence: a note on the physical backgrounds of two types of light microscopy.

Bright microscopic images against a dark background can be originating not only from fluorescence, but also from selective reflection. Selective reflection or scattering of visible light in microscopic preparations can be used for the visualization of sometimes otherwise barely distinguishable material. The images obtained superficially resemble those from fluorescence microscopy. They do not, however, result from huminescence but from selectively reflected light with wavelengths in the region of the absorbance peak of the chromophore present in the stained biological material. The respective backgrounds of the underlying physical phenomena and the conditions under which selective reflection can occur are discussed.     

The effect of environmental enrichment on the behaviour of caged rabbits (Oryctolagus cuniculus)

Behaviour and use of the cage area were studied in 96 rabbits (Oryctolagus cuniculus) kept in an enriched cage system - with access to shelter and raised height at the back of the cage - and in a conventional cage system to estimate the effects of the environmental enrichment on the rabbits' welfare. The rabbits' behaviour and placement in the cage were observed, using continuous video recording through 24 h and direct scan sampling during the daytime. In addition, an open-field test was carried out with each rabbit, and after every single test, the rabbits' timidity of being captured was recorded. Rabbits kept in the conventional cage system, especially the females, showed more restlessness, excessive grooming, bar-gnawing and timidity than rabbits kept in the enriched cage system. This indicates increased stress in the rabbits kept in the conventional cage system. All the rabbits performed most of the active behavioural elements in the daytime and were resting mostly at night that shows that the rabbits in both cage systems were adapted to the daily activity in the animal unit; the enrichment had no effect on the daytime activity.Only a few rabbits, particularly the females, used the box as a shelter or resting-place. On the other hand, they more often used the roof of the box as a look-out or resting-place. Furthermore, the rabbits' behaviour showed that they utilised the raised height in the enriched cage system.These results indicate that rabbits kept in an enriched cage system, particularly the females, had better welfare than rabbits kept in a conventional cage system because they had access to shelter and a better chance of interacting with the environment.     

Behaviour of 3-week weaned pigs in Straw-Flow(R), deep straw and flatdeck housing systems

The behaviour of 3-week weaned pigs in different housing systems was examined as part of an assessment of the suitability of the Straw-Flow(R) system for pigs of this age. Three replicate pens of 20 pigs were weaned at 6.4 kg liveweight into each of: (a) deep-straw; (b) Straw-Flow(R); (c) large flatdeck; (d) small flatdeck. A kenneled lying area was provided in (a) and (b). The floor in (c) and (d) was expanded metal. Stocking densities were 0.23 m(2)/pig in (a), (b) and (c), and 0.17 m(2)/pig in (d). After 4-5 weeks (at 19.6 kg liveweight), 16 pigs from each pen were moved into Straw-Flow(R) grower pens (0.68 m(2)/pig) and observed until slaughter at 90.6 kg. Pigs in systems incorporating straw showed behaviour patterns associated with increased welfare (greater straw-directed behaviour and less pig-directed and pen-directed behaviour) relative to those in barren pens. Behavioural differences between (a) and (b) related to differences in available straw; there were few differences between (c) and (d). Pigs from (c) and (d) showed increased rooting relative to those from (a) after transfer to the grower pens, but other behavioural differences between weaner treatments did not persist. It is concluded that the Straw-Flow(R) system can provide suitable accommodation for weaned pigs.     

Affinity enhancement of bispecific antibody against two different epitopes in the same antigen.

For the enhancement of antibody binding affinity, a bispecific antibody against two different epitopes in human chorionic gonadotropin hormone, one is in alpha-subunit and the other is in beta-subunit, was prepared by chemical recombination using 5,5'-dithiobis(2-nitrobenzoic acid). The epitopes recognized by antibodies were investigated by competitive radioimmunoassay, two-site sandwich radioimmunoassay and additivity assay and a proper epitope pair was chosen for preparation of the bispecific antibody. This bispecific antibody has dual specificity and as much as 17.2-fold higher affinity than that of monoclonal antibody with higher affinity by dual antigen binding radioimmunoassay and Scatchard plot analysis.     

Mechanism of photoinactivation in photosynthetic systems II. The occurrence and properties of two different types of photoinactivation

Photoinactivation of the activity of NADP photoreduction withreduced DPIP or with reduced TMPD as the electron donor wasinhibited by the absence of oxygen in the atmosphere or by thepresence of photosynthetic inhibitors (CMU, DCMU, o-phenanthroline)in the preillumination mixture. Photoinactivation of the photoreductionof NADP or DPIP with water as the electron donor was not affected,or even accelerated, by these conditions of preillumination.The concentrations of inhibitors required for maximum inhibitionin the former case corresponded to those required for inhibitionof photosynthetic electron transport. The results indicatedthe occurrence of 2 different types of photoinactivation, eachspecifically affecting photosystems I and II, and differingin behaviours; including their requirement for oxygen in theatmosphere and their responses toward the presence of photosyntheticinhibitors during the preillumination period. (Received July 30, 1969; )     

Organic calcium channel blockers generate the coexistence of two different types of action potentials in the same muscle membrane following chemical induction of excitability.

1. Following exposure to the sulfhydryl reagents known as alpha,beta-unsaturated carbonyl compounds, the ventroabdominal flexor muscles of the crustacean Atya lanipes, which are normally completely inexcitable, generate trains of overshooting calcium action potentials; the effects of organic calcium channel antagonists and potassium channel blockers on the chemically-induced trains of action potentials have been studied. 2. Verapamil and D600, at micromolar concentrations, elicit the appearance of slow, cardiac-like action potentials which coexist with the much faster chemically-induced calcium spikes, transforming the regular repetitive firing into a cyclic bursting pattern. 3. Bepridil (1 microM) decreases the frequency of firing of the action potentials, probably by increasing the threshold for the activation of a population of the chemically-induced calcium channels. 4. The potassium channel blockers, TEA (30-40 mM) and quinidine (100-200 microM), delayed the rate of repolarization of the chemically-induced action potentials; none of the potassium channel blockers, however, induced the appearance of repetitive spike activity.