Assessing the Transfer of Genetically Modified DNA from Feed to Animal Tissues

In Europe, public and scientific concerns about the environmental and food safety of GM (Genetically Modified) crops overshadow the potential benefits offered by crop biotechnology to improve food quality. One of the concerns regarding the use of GM food in human and animal nutrition is the effect that newly introduced sequences may have on the organism. In this paper, we assess the potential transfer of diet-derived DNA to animal tissues after consumption of GM plants. Blood, spleen, liver, kidney and muscle tissues from piglets fed for 35 days with diets containing either GM (MON810) or a conventional maize were investigated for the presence of plant DNA. Only fragments of specific maize genes (Zein, Sh-2) could be detected with different frequencies in all the examined tissues except muscle. A small fragment of the Cry1A(b) transgene was detected in blood, liver, spleen and kidney of the animals raised with the transgenic feed. The intact Cry1A(b) gene or its minimal functional unit were never detected. Statistical analysis of the results showed no difference in recovery of positives for the presence of plant DNA between animals raised with the transgenic feed and animals raised with the conventional feed, indicating that DNA transfer may occur independently from the source and the type of the gene. From the data obtained, we consider it unlikely that the occurrence of genetic transfer associated with GM plants is higher than that from conventional plants.     

Evaluation of Microcystis as food for zooplankton in a eutrophic lake

Four experiments were conducted to evaluate Microcystis as food for zooplankton in Lake Kasumigaura, and the following results were obtained. (1) Moina micrura (Cladocera) showed little growth and no reproduction when the animal was reared with Microcystis cultured in the laboratory. The animal did not grow nor reproduce well when Chlorella was mixed with Microcystis as food. (2) Moina micrura assimilated Microcystis much less than Chlorella when the animal fed on single species of Microcystis or a mixture with Chlorella. (3) Microcystis collected from Lake Kasumigaura could not be utilized by Moina micrura even though the colonies were broken up into edible sizes. However, the alga turned into utilizable food when it was decomposed. (4) No inhibitors of Moina micrura population growth could be found in the non-filtered water of Lake Kasumigaura where Microcystis was blooming heavily. Decomposed Microcystis seemed to be utilized by zooplankton as an important food source in Lake Kasumigaura.     

Laboratory study of the grass shrimp feeding preferences

Grass shrimps are very common in various aquatic habitats, and are generally considered omnivorous. But, despite the numerous studies on interspecific trophic interactions, few studies have evaluated their feeding strategy, especially by experimental approaches.In this study, we determined, in the absence of both intra and interspecific competitive interactions, the feeding preferences of the freshwater grass shrimp Palaemonetes antennarius for the most common potential resources in its habitat. Plant and animal items were offered as food to each adult shrimp in multiple-choice laboratory experiments. Resource selection was determined under conditions of low and high food availability and related to the shrimp body size. The role of microbial colonisation in favouring detritivory was also investigated.Results of these experiments showed that P. antennarius consumed only animal items, mostly preferring the isopod Proasellus coxalis. In addition, palatability of the unselected plant remains and non-living prey was not improved by the microbial enrichment. Shrimps also resorted to cannibalism at low ration. By contrast, at high ration they specialised on a narrower range of resources, directing more their feeding efforts towards isopods. The preference ranking of the selected food types was inversely related to the average mass of the prey and varied with the shrimp body size. In particular, the number of live isopods in the diet increased exponentially with the shrimp mass at food high ration, which resulted not to be limiting. In conclusion, food availability influences the food niche of P. antennarius whose feeding strategy in the laboratory is facultative specialisation on dead and live animals rather than omnivory. Ease of handling and/or capture seems to play a role in the food preference ranking.     

The association between the phytoplankton, Rhopalosolen species (Chlorophyta; Chlorophyceae), and Anopheles gambiae sensu lato (Diptera: Culicidae) larval abundance in western Kenya

Algae are important food resources of the larvae of the African malaria vectors, Anopheles gambiae Giles and Anopheles arabiensis Patton (Anopheles gambiae sensu lato), and other zooplankton, but empirical evidence remains meager about the agal flora in ephemeral water bodies. The animals present in natural aquatic habitats in western Kenya were sampled from July to November 2002 to study abiotic and biotic environmental factors determining A. gambiae sl larval abundance. The five highest concentrations of third and fourth instars and pupae (hereafter referred to as old-stage larvae) were sampled in conjunction with the unicellular epizoic green algae, Rhopalosolen species (Chlorophyta; Chlorophyceae). Canonical correspondence analysis revealed that the presence of Rhopalosolen species was the most important determinant of the animal assemblage. The density of old-stage A. gambiae sl larvae was positively correlated with the presence of Rhopalosolen species, but the density of first and second instars of A. gambiae sl was not. The water bodies with Rhopalosolen sp. yielded larger mosquitoes in spite of the higher density of larvae. We demonstrated that the productivity of water bodies in terms of the larvae of malaria vectors can differ in magnitude depending on the agal flora. We discuss phytoplankton as a regulator of mosquito larval populations.     

Understanding the threats to biological diversity in southeastern Brazil

Habitat destruction and the over-exploitation of species are perhaps thetwo most important processes threatening biodiversity. Whilst the growing humanpopulation puts considerable pressure on biological resources, a number ofsocial and economic factors tend to augment over-exploitation of theseresources. Here we show that the over-exploitation of the tropical palm treeEuterpe edulis Mart., as a consequence of social problems,has contributed significantly to its extinction in several forest fragments andalso to the disappearance of many wild animal species which used to be found inareas of the Atlantic Rain Forest (Brazil). Some of these species have been lostas a direct result of people hunting the animals when they went harvestingE. edulis while others disappeared as a consequence of thedecrease in food availability or by alterations in the trophic structure.     

Pneumocystis carinii: Genetic Diversity and Cell Biology

As an important opportunistic pulmonary pathogen, Pneumocystis carinii has been the focus of extensive research over the decades. The use of laboratory animal models has permitted a detailed understanding of the host–parasite interaction but an understanding of the basic biology of P. carinii has lagged due in large part to the inability of the organism to grow well in culture and to the lack of a tractable genetic system. Molecular techniques have demonstrated extensive heterogeneity among P. carinii organisms isolated from different host species. Characterization of the genes and genomes of the Pneumocystis family has supported the notion that the family comprises different species rather than strains within the genus Pneumocystis and contributed to the understanding of the pathophysiology of infection. Many of the technical obstacles in the study of the organisms have been overcome in the past decade and the pace of research into the basic biology of the organism has accelerated. Biochemical pathways have been inferred from the presence of key enzyme activities or gene sequences, and attempts to dissect cellular pathways have been initiated. The Pneumocystis genome project promises to be a rich source of information with regard to the functional activity of the organism and the presence of specific biochemical pathways. These advances in our understanding of the biology of this organism should provide for future studies leading to the control of this opportunistic pathogen.     

Comparison of pressure chamber and psychrometer estimates of leaf water potential in rice

Summary The need to compare pressure-chamber estimates of leaf water potential with a psychrometric method has been established for several crop species. We investigated this relationship for rice (Oryza sativa L.) as well as the need to protect leaves from water loss during sampling and measuring period in the pressure chamber. Two rice cultivars grown in containers on a clay-loam soil were stressed to varying degrees by withholding water. Fully expanded leaves were sampled for estimation of leaf water potential by the dew point hygrometer and pressure-chamber techniques. The same leaf was used in both methods allowing direct comparison. Additionally, two alternative methods of leaf handling for measurement by the pressure chamber technique were compared. Protection of leaf samples against water loss during excision, transport and handling was found to be more important at higher leaf water potentials (>−1.0 MPa). The two cultivars used appeared to differ in their response to protection of the leaf sample. These results serve to further caution pressure chamber users on extrapolating comparisons between the two measurement methods and between tissue handling techniques even within a crop species.     

Handling and Welfare of Bovine Livestock at Local Abattoirs in Bangladesh

The World Organization for Animal Health (OIE) allows rope casting and the tying of legs for nonhuman animal slaughter without stunning. The handling and welfare of bovine livestock (Bos indicus and Bubalus bubalis) were studied in 8 local abattoirs in 5 districts of Bangladesh. A total of 302 animals were evaluated. At the local abattoirs, approximately 1/3 of the cattle and water buffalo were either emaciated or injured/sick. The size and vigor of the animals determined the casting method. Small and weak animals were cast on concrete floors by lifting a foreleg followed by pushing, or simply by twisting the head of the animal and then binding the legs with rope. Vigorous animals such as buffalo were cast using ropes and human force. Bleeding was slow and flaying was sometimes initiated before the animals were unconscious. Pulling and tearing of the trachea and pouring of water into the exposed trachea shortly after cutting were also observed in some cases. The overall animal handling was unnecessarily rough and the OIE standards were not implemented. Animals are subjected to considerable mistreatment, and there is an urgent need for the training and education of the staff in abattoirs concerning humane slaughtering practices as well as a need to build modern slaughtering plants in Bangladesh.     

Diet-dependent depletion of queuosine in tRNAs in Caenorhabditis elegans does not lead to a developmental block

Queuosine (Q), a hypermodified nucleoside, occurs at the wobble position of transfer RNAs (tRNAs) with GUN anticodons. In eubacteria, absence of Q affects messenger RNA (mRNA) translation and reduces the virulence of certain pathogenic strains. In animal cells, changes in the abundance of Q have been shown to correlate with diverse phenomena including stress tolerance, cell proliferation and tumour growth but the function of Q in animals is poorly understood. Animals are thought to obtain Q (or its analogues) as a micronutrient from dietary sources such as gut microflora. However, the difficulty of maintaining animals under bacteria-free conditions on Q-deficient diets has severely hampered the study of Q metabolism and function in animals. In this study, we show that as in higher animals, tRNAs in the nematode Caenorhabditis elegans are modified by Q and its sugar derivatives. When the worms were fed on Q-deficient Escherichia coli, Q modification was absent from the worm tRNAs suggesting that C. elegans lacks a de novo pathway of Q biosynthesis. The inherent advantages of C. elegans as a model organism, and the simplicity of conferring a Q-deficient phenotype on it make it an ideal system to investigate the function of Q modification in tRNA.     

Behavioural and physiological stress responses to handling in wild guanacos

Both ethology and physiology can provide insights about stress conditions in wild animals during handling. Lama guanicoe (guanaco) is the major large herbivore inhabiting arid zones in Argentina. Since 2005, the Payún Matrú Cooperative, a business established and run by native people, has captured and handled guanacos for shearing at Payunia Reserve, Mendoza Province, Argentina, to market the species’ fibre for clothing and other products. The aim of our study was to evaluate the behavioural and physiological stress responses to handling of wild guanacos during these shearing activities. We evaluated stress conditions in guanacos by monitoring the frequency of agonistic behaviour in the holding corrals, following capture, the rate of vocalisation and agonistic behaviour during handling, and by estimating cortisol levels in handled animals. Crowding increased agonistic behaviour; this appeared to be related to dominance conflicts between males in the corrals. Time of corralling increased rate of agonistic behaviour and vocalisations during handling. The rate of behavioural stress responses was negatively related to cortisol levels in males; which may be due to animal fatigue or because behavioural stress responses may help mitigate physiological stress responses. Incorporating our results into animal welfare practices could help reduce stress conditions in wild guanacos. Our results suggest that both behavioural and physiological responses are important measures of stress in handled animals, and that behavioural stress responses cannot be used as a simple surrogate for glucocorticoid levels.     

Sexual reproduction in Daphnia: interspecific differences in a hybrid species complex

To investigate whether reproductive variation in the Daphnia galeata-cucullata-hyalina hybrid species complex can explain the occurrence of hybrids and backcrosses, a total of 43 clones from three north German lakes were tested for allocation to sexual reproduction under equal stress conditions. Six replicates per clone were followed until the seventh adult instar. I used the following cues to promote sexual reproduction: short photoperiod, water from a crowded Daphnia culture, fish-conditioned water and low food concentration. For each animal, clutch size and clutch sex were recorded. Ephippia, which were empty since the animals were cultured individually, were only produced by D. cucullata and D. cucullata x hyalina (26% and 6% of the broods, respectively), whereas almost all taxa produced males (the range was 2–15%). Intraspecific variation for male and ephippial production was also found. The fact that the taxa tested show different responses to the stimuli of-fered may indicate that there is reproductive isolation between them. This is supported by field data from the Tjeukemeer (The Netherlands) which shows that D. galeata mostly produces ephippia in spring and D. cucullata in autumn. The presence of hybrids with ephippia in both seasons, however, shows that backcrossing is also possible.     

Effect of physiological state and fungal inoculation on chemically modulated host-plant finding by Carpophilus hemipterus and Carpophilus lugubris

Host orientation by Carpophilus hemipterus L. and Carpophilus lugubris Murray (Coleoptera: Nitidulidae) was investigated in a horizontal wind tunnel to ascertain if these species differ in their response to aseptic and fungal-inoculated hosts, and also to determine how age, nutritional status, diel period, and locomotory opportunity affect these behavioral responses. Both species responded to food odors beginning on Day 3 of adult life by walking upwind to the source; flight activity and flights to the odor source began for C. hemipterus on Day 4, but C. lugubris continued to walk to the source and rarely flew regardless of age. Both species displayed maximum response to food odors from 6 to 9 days after emergence and showed bimodality in host orientation during the photophase. C. hemipterus maintained with artificial diet took flight as frequently as when maintained with water only, as long as they were deprived of diet for 36 h before the test; however, after takeoff, beetles maintained with diet were less likely to fly to food odor in comparison to beetles maintained with water. C. lugubris maintained with diet until 36 h before the test displayed a reduction in walks upwind to sources of food odor compared to beetles maintained with water. Three hours of unrestricted locomotion under a high-pressure sodium lamp did not enhance upwind orientation to host odors by C. hemipterus maintained with diet or water, or by C. lugubris maintained with water; however, such preexposure to a sodium lamp resulted in reduced takeoffs in C. hemipterus maintained with artificial diet. Both species were attracted to all fruit and vegetable substrates offered; however, aseptic substrates were less attractive than were substrates inoculated with Saccharomyces cerevisiae Hansen or Candida krusei (Castellani) Berkhout for C. lugubris. Despite the broad host range exhibited by these two nitidulid species, each responded to chemical cues from longrange (2.5 m), a trait once assigned to specialists.     

A history of recent advancements on <Emphasis Type="Italic">Nephrops norvegicus</Emphasis> behavioral and physiological rhythms

Biological rhythms are a widespread feature of living organisms, being expressed at any level of their organization. Behavioral and physiological rhythms can affect the results on species stock assessment when the timing of sampling is not taken into account. That timing is of importance since animals may be present or not in a certain area of sampling depending on their activity cycle. As an example of this, the rhythmic behavior and physiology of one of the most commercially important European decapods, the Norway lobster (Nephrops norvegicus) was studied. These rhythms affect its commercial catchability at a diel and at a seasonal scale. Nephrops inhabits muddy bottoms where animals dig burrows that save them from the trawl tow capture when occupied. Catch patterns have been widely used as proxy of activity rhythms of populations of different depths. Catches show a modulation upon the day-night cycle since animals emerge under optimum environmental illumination in order to feed. Emergence is also affected in a not fully clarified manner by other variables of environmental and demographic nature (e.g. food presence, hunger state, sex, size, reproductive stage, territorialism and mating). All these features make Nephrops a good model of reference for studies on biological rhythms of other commercially important deep water decapods in relation to their ecological context. In this review, we summarize the actual knowledge on Nephrops behavioral and physiological rhythms. We will compare data obtained from laboratory tests on single individuals with data obtained from trawling of populations of different depths. We will also describe some new hypotheses on the rhythmic regulation of the species behaviour, as well as potential scenarios for future research.     

Stress resistance induced by paramylon treatment in Artemia sp.

A study was undertaken to investigate the effectiveness of paramylon extracted from the non-photosynthetic WZSL mutant of Euglena gracilis in potentiating the resistance of the brine shrimp Artemia sp. to stress conditions resulting from poor growth medium quality and daily handling. The experimental design incorporated four different treatments: I) paramylon addition/no growth medium replacement; II) no paramylon addition/no growth medium replacement; III) paramylon addition/growth medium replacement; IV) noparamylon addition/growth medium replacement. As shown by the survival curves of Artemia sp., the addition of paramylon significantly reduced susceptibility of animals to the stressors. Paramylon effectively increased also the ability of offspring to withstand poor water quality and handling damages. Without paramylon almost all offspring died before adulthood, whereas addition of paramylon allowed the animals to moult to the adult stage. Moreover, reproductive success (higher number of living offspring) was enhanced considerably in animals treated with paramylon treated under both stress conditions. These results show that paramylon extracted from Euglena represents a promising biologically active compound for aquaculture purposes. It could be used as a purified product or as component of whole cells, since the Euglena mutant, because of the high intracellular amount of paramylon it can accumulate, could be added to the feed or to water in tanks and ponds without prior processing. This revised version was published online in August 2006 with corrections to the Cover Date.     

Effects of recent experience on foraging in tephritid fruit flies

In field-cage studies, we investigated how the foraging behavior of tephritid fruit flies is modified by experience immediately prior to release on host plants. We observed females of a relatively monophagous species,Rhagoletis mendax (blueberry maggot fly), an oligophagous species,Rhagoletis pomomella (apple maggot fly), and a polyphagous species,Ceratitis capitata (Mediterranean fruit fly). Just prior to release on a host plant, the following kinds of stimuli were supplied: (1) single oviposition in a host fruit, (2) contact with 20% sucrose, (3) contact with a mixture of protein food (bird feces and sucrose), (4) contact with water, and (5) a walk over a host-plant leaf. When flies foraged on host plants without resources, search was most intensive (as measured by number of leaves visited) following a single oviposition in fruit, but residence time generally was the same following exposure to sugar, protein, and fruit stimuli.Rhagoletis mendax andC. capitata females visited the fewest leaves following exposure to water or host leaves, whereasR. pomonella foraged equally intensively following exposure to food stimuli, water, or leaves. On host plants containing resources (fruit and protein food), a single oviposition dramatically increased the number of females of all three species that found fruit compared to females that received experience with food, water or foliar stimuli. We found no significant effect of recent brief experience with any of the stimuli on subsequent attraction to protein food. Overall,C. capitata exhibited a higher propensity to abandon host plants than eitherR. mendax orR. pomonella. We suggest that this may reflect adaptations to differences in distribution of host plants in nature, strategies of dispersal, and host range.     

Home Range,Population Density,and Food Resources of Agouti paca (Rodentia: Agoutidae) in Costa Rica: A Study Using Alternative Methods1

Field studies of elusive mammals such as Agouti paca are difficult, and results are often biased and incomplete because the animals are nocturnal and avoid traps. By studying an A. paca population in a Pacific lowland forest of Costa Rica, we developed indirect observational methods to estimate population density and food resource utilization. To estimate population density, we divided the number of A. paca burrows found in a 12-ha census area by the average number of burrows utilized per individual from radiotelemetry data. We compared this estimate to independent population estimates obtained using two transect methods. Agouti paca density was estimated at 93 individuals/km² based on burrow number, and between 67 to 70 individuals/km² using transect methods. This discrepancy corroborates previous studies suggesting that transect methods underestimate population density. To identify food resources consumed by the species, we collected fruits and seeds with unambiguous A. paca teeth marks. The only animal in Central America with an incisor width similar to A. paca. is Dasyprocta variegata; in a comparison of both species, however, we found that A. paca had a significantly larger incisor width and that teeth marks of 4 mm or wider unambiguously indicated handling by this species. By applying this technique, food resources utilized by A. paca can be identified more easily than by direct observation. At our study site, we identified 33 plant species consumed by A. paca; 61 percent are new records for this animal. Our results suggest that A. paca plays a major, although not fully understood, ecological role in seed predation and dispersal.     

Hand preference study in marmosets (Callithrix jacchus) using food reaching tests

Hand preference has been investigated in New World primates but the data obtained thus far are controversial. In this study we investigated hand preference in common marmosets,Callithrix jacchus, during the execution of a reaching for food task. We used 46 adult common marmoset males (n=27) and females (n=19) from the Universidade of Rio Grande do Norte colony, both wild and captive-born. To test the hand preference we used a device measuring 10 cm², with a central hole 1 cm in diameter, to force the animal to use only one hand to reach for food on a food dish located underneath. Each animal was tested 5 times and had to make a maximum of 20 successful attempts per session. A total of 100 successful attempts per animal and 4,600 successful attempts for all animals were recorded during the experiment. Latency and duration of the sessions were measured and we found preference for the use of one of the hands in common marmoset individuals, i.e. 45 of total of 46 animals used significantly more the right or the left hand when performing the task. However no bias at the population level was found. Females born in captivity presented an increase in the duration of latency for the first successful attempt and in the total duration of the test sessions. These findings might be indicating differences associated with a natural tendency for females to be more selective and to spend more time exploring alimentary sources. Additionally, captive-born females may have a constrain in developing cognitive abilities regarding foraging since they have food available during most part of the time.     

Experimental study of vibrio parahaemolyticus (biotype 2) transfer from water and sediments to benthic marine food chain organisms]

Transfer of Vibrio parahaemolyticus (biotype 2) from sediments to water and from water to benthic marine organisms was studied experimentally using a streptomycin-resistant strain. Transmission by trophic pathways was also studied using reconstituted marine food chains (Mytilus edulis, Nereis diversicolor, Carcinus maenas, Scorpaena porcus, Mus musculus). Water colonization by sediments could be observed only at temperatures above 16 degrees C. Sediments could well constitute a disseminating reservoir for these germs, their cycle in water being dependent of the cycle followed in the sediments. Contamination of animal organisms is essentially effected by a direct mean, either water or sediments; transfer by trophic pathways being negligible. Infection of land consumers (mice) is linked quantitatively to the nature of the last marine organism of the food chain since bacteria can flourish in the digestive tract of certain animals (Carcinus maenas).     

Isolation and physiological characterisation of Thiobacillus thyasiris sp. nov., a novel marine facultative autotroph and the putative symbiont of Thyasira flexuosa

A novel facultatively chemolithoautotropic Thiobacillus, isolated from the gill tissue of the marine bivalve Thyasira flexuosa, is described. It is believed to be the symbiont from this animal, providing the animal with carbon fixed by the Calvin cycle. The organism grows lithoautotrophically on thiosulphate, tetrathionate and elemental sulphur, which are oxidised to sulphate. It oxidizes sulphide, thiosulphate, trithionate, tetrathionate and hexathionate, but not thiocyanate. Kinetic constants for these substrates are presented. In autotrophic batch culture it produces yields that are among the lowest reported for thiosulphate or tetrathionate as energy substrates (1.25 and 2.5 g cell-carbon per mol substrate, respectively). Autotrophic cultures contain ribulose bisphosphate carboxylase and excreted 20% of their fixed carbon into the medium during growth. Mixotrophic growth on acetate and thiosulphate resulted in partial repression of the carboxylase. The organism is slightly halophilic and markedly halotolerant, showing optimum growth at about pH 7.5 and maximum growth rate at 37° C. It contains ubiquinone Q-10 and its DNA contains 52 mol % G+C. These characteristics distinguish it from any other Thiobacillus or Thiomicrospira species previously described. The organism is formally described and named as Thiobacillus thyasiris.     

Dynamics of incisor growth and daily increments on the incisor surface in three species of small rodents

The lower and upper incisors has been studied in striped field mice (Apodemus agrarius), yellow-necked mice (A. flavicollis), and migratory hamsters (Cricetulus migratorius) trapped in the Volgograd Region from April to March. The incisors of all animals have been found to have a striated surface, with series of slight ridges and grooves. As shown by analyzing the incisors of animals labeled with tetracycline, these are daily increments of dentin. The total numbers of increments (the period of complete tooth renewal) in the upper and lower incisors of the same animal are similar, but the degrees of their distinctness can be different. The number of daily increments increases but their width (daily growth rate) decreases with age even in adult animals and, in addition, varies by seasons. These two trends—the decrease in tooth increment with age and its seasonal variation—have proved to mask each other when an uneven-aged sample of animals trapped over several months is studied. Therefore, the season of death of an animal cannot be reliably estimated from the width of daily tooth increment unless the age of this animal is taken into account. The increment width in young animals in autumn may be the same as in older animals in spring and summer. In attempts to use the incisor surface sculpture for determining the season of animal death, it is necessary to separate the young from adults (at least by the criterion of incisor length). The decrease of increment width with age and its seasonal variation in the absence of obvious changes in the diet and pattern of incisor attrition indicate that the incisor growth rate is subject to age-dependent and seasonal changes, as is the growth rate of the organism as a whole. The distinctness and internal structure of daily increments also vary depending on animal age and species, which is due apparently to differences in the circadian rhythms of incisor growth. The presence of one or, less frequently, several grooves within a daily increment, which has occurred in all species studied, is evidence for not only circadian but also ultradian rhythms in the growth of incisors.