A pan-neotropical analysis of hunting preferences.

Hunting in the neotropics is a widespread form of resource extraction. However, there is increasing concern that current activities are leading to the decline and extirpation of vulnerable species; particulary ateline primates, large ungulates (such as tapirs and white-lipped peccaries) and large birds such as curassows. Hunting patterns are expected to be a product of two principal influences: the value of return for a given amount of effort invested into hunting, and cultural factors that determine the prestige and usefulness of prey. Previous work has suggested that hunting profiles change in a predictable way over time, becoming more diverse and more dependent on smaller bodied species as preferred, large-bodied prey become scarcer. In this paper, we evaluate the hunting profiles of 78 neotropical communities in Central and South America. We investigate the uniformity of species preferences, whether communities that are geographically closer have similar hunting profiles, and whether the age and size of settlements can be used to predict the type and diversity of species targeted. We found that there was only a weak correlation between the structure of communities' hunting profiles and their geographical proximity. Neither a community's size nor age was a good predictor of the shape and structure of its hunting profile. Our data suggest that either the availability of prey or the cultural influences dictating the value of different species can change rapidly over small distances, and that older and larger settlements do not impact prey species distributions in a predictable way.

Impacts of UVB provision and dietary calcium content on serum vitamin D3 , growth rates, skeletal structure and coloration in captive oriental fire-bellied toads (Bombina orientalis).

Many amphibian species are dependent on ex situ conservation interventions for their long-term persistence. However, projects have been jeopardised by husbandry issues involving poor calcium metabolism and nutritional metabolic bone disease (NMBD). Healthy calcium metabolism requires appropriate dietary calcium content and access to vitamin D3 . In many animals, vitamin D3 can be photobiosynthesised in skin exposed to UVB radiation, as well as extracted from the diet, but the extent of vitamin D3 photobiosynthesis in amphibians is poorly known. Additionally, prey insects for captive amphibians are deficient in calcium and calcium content must be artificially increased, but the effects of different levels of augmentation and their interaction with UVB exposure are also little understood. We fed captive fire-bellied toads (Bombina orientalis) with crickets augmented to contain 5% and 10% calcium and housed them with and without UVB exposure. Despite additional dietary vitamin D3 supplementation, we found that toads exposed to UVB radiation exhibited significantly higher serum vitamin D3 levels, indicating that this species may partly rely on photobiosynthesis sources of vitamin D3 . These data are the first to show a direct link between UVB exposure and serum vitamin D3 in an amphibian. We found significant positive effects of UVB exposure and 10% dietary calcium content on skeletal structure, as well as complex interactions between treatments. We also found UVB radiation exposure resulted in more rapid natural coloration acquisition. Together, this indicates that standard calcium plus vitamin D3 supplementation methods may not fully substitute for UVB exposure and for increased feeder insect calcium content. This may have implications for the success of ex situ amphibian conservation, as well as for the welfare of captive amphibians in general. Our data lend support for the provision of UVB radiation for captive, basking amphibians.

Virological failure at one year in triple-class experienced patients switching to raltegravir-based regimens is not predicted by baseline factors.

We evaluated rates and determinants of virological failure in triple-class experienced patients receiving raltegravir-based regimens from a national observational study over 48 weeks, defined by any one of the following: (1) no HIV-RNA suppression to undetectable levels (<50 copies/mL) during follow-up; (2) detectable viral load after obtaining undetectable levels; and (3) leaving the study before 48 weeks. Among 101 eligible patients, 26 (25.7%; 95% CI 17.2-34.2) had virological failure. No significant differences between patients with and without virological failure were observed for gender, age, route of transmission, baseline CD4/HIV-RNA, CDC group, hepatitis B or C co-infections, resistance (based on the last genotype available), type and number of concomitant drug classes, concomitant use of darunavir, atazanavir, etravirine, enfuvirtide or maraviroc, and health-related quality-of-life measures. A high rate of treatment response was observed. The analyses did not identify any baseline factor associated with failure, including resistance status. Even if we cannot exclude the presence of pre-existing minority resistant variants not captured by genotypic tests, the lack of baseline predictors of failure suggests the need to monitor patients closely during follow up for other factors, such as potential drug interactions and reduced levels of adherence, which may favour virological failure.

Can carotenoids mediate the potentially harmful effects of ultraviolet light in Silurana (Xenopus) tropicalis larvae?

Amphibians have recently experienced unprecedented declines in the wild, the causes of which are often difficult to mitigate. This has increased the importance of ex situ conservation; however, long-term maintenance and breeding of amphibians in captivity often has limited success. In vertebrates, vitamin D is required for calcium homeostasis and is produced endogenously in skin exposed to ultraviolet (UV) light; however, UV light can be harmful to amphibians in some instances. Carotenoids are dietary pigments that may provide protection from UV light. The aim of this study was to assess the protective capability of carotenoids against the potentially harmful effects of UV light in Silurana (=Xenopus) tropicalis larvae raised in an enhanced or limited UV environment. Tadpole survival and the size and developmental stage reached by the end of the study period were measured. Carotenoids had a significantly positive effect on developmental rate in both UV-limited and UV-enhanced environments. Larvae in an enhanced UV environment were significantly larger than those raised under a limited UV environment, irrespective of diet. Carotenoid-fed larvae tended to have increased survival in relation to those raised without carotenoids, but only in a limited UV environment. Carotenoids appear to provide little protection against UV light in this case. The role of carotenoids in amphibian health has not previously been studied. We show that carotenoid availability significantly influences development and may increase survival in S. (X.) tropicalis larvae. This finding may have important implications for recommendations made on the nutrition of amphibians in captivity.

Quantitative trait loci mapping of phenotypic plasticity and genotype-environment interactions in plant and insect performance.

Community genetic studies generally ignore the plasticity of the functional traits through which the effect is passed from individuals to the associated community. However, the ability of organisms to be phenotypically plastic allows them to rapidly adapt to changing environments and plasticity is commonly observed across all taxa. Owing to the fitness benefits of phenotypic plasticity, evolutionary biologists are interested in its genetic basis, which could explain how phenotypic plasticity is involved in the evolution of species interactions. Two current ideas exist: (i) phenotypic plasticity is caused by environmentally sensitive loci associated with a phenotype; (ii) phenotypic plasticity is caused by regulatory genes that simply influence the plasticity of a phenotype. Here, we designed a quantitative trait loci (QTL) mapping experiment to locate QTL on the barley genome associated with barley performance when the environment varies in the presence of aphids, and the composition of the rhizosphere. We simultaneously mapped aphid performance across variable rhizosphere environments. We mapped main effects, QTL × environment interaction (QTL×E), and phenotypic plasticity (measured as the difference in mean trait values) for barley and aphid performance onto the barley genome using an interval mapping procedure. We found that QTL associated with phenotypic plasticity were co-located with main effect QTL and QTL×E. We also located phenotypic plasticity QTL that were located separately from main effect QTL. These results support both of the current ideas of how phenotypic plasticity is genetically based and provide an initial insight into the functional genetic basis of how phenotypically plastic traits may still be important sources of community genetic effects.

Plant genotype mediates the effects of nutrients on aphids.

Soil nutrients, and factors which influence their concentrations and bioavailability, form a basic component of bottom-up control of ecosystem processes, including plant-herbivore interactions. Increased nutrient levels are linked, through plant defence theory, with increased levels of herbivore susceptibility. The focal point of many ecological experiments examining this link is at the species level, where the response of single species is the average of many different genotypes. Here, we focus on the genetic basis of indirect ecological interactions. We investigated the effects of nutrient concentration on the population growth of an aphid herbivore across multiple genotypes of barley in relation to plant growth rate. In general, both aphid population size and plant growth rate increased with nutrient concentration. However, they were both dependent on the interaction between nutrient concentration and barley genotype. Our data raise the testable possibility of differential defense responses between genotypes of barley, for example divergent, fixed and inducible defences against aphids. We provide evidence that the indirect effects of soil nutrients on aphid population size are mediated by the genetics of the host plant.

Tritrophic effects on survival and development in a Coccidophagous coccinellid.

Chilocorus nigritus is currently considered one of the most successful biological control agents of armoured scale insects. However, establishment of this beetle in crop pest situations has not always been successful and there are still gaps in our knowledge of its ecology and behaviour. The research involved an examination of tritrophic effects on the survival and development of this common diaspid predator. The effect of a forced change in host plant on the developmental time of the juvenile stages was also examined. The prey and host plants used were the armoured scales Aspidiotus nerii Bouché Homoptera: Diaspididae and Abgrallaspis cyanophylli (Signoret) Homoptera: Diaspididae, on potatoes (Solanum tuberosum L.) and Butternut squashes (Cucurbita moschata Duchesne ex Lamarck). C. nigritus eggs were incubated on four treatments of scales on potatoes or squashes for ten days, half the second instar larvae were then switched to the same scales on the other host plant. Daily observations were made during development to adult emergence. C. nigritus larvae survived and completed development on two species of diaspid scales and the two host plants examined with varying levels of success. Larvae were able to switch from feeding on A. nerii on potatoes to A. nerii on squashes or A. cyanophylli on potatoes to A. cyanophylli on squashes and vice versa with little or no deleterious effects when compared to those beetles reared on one prey and host plant throughout. There were significant differences in survival of larvae reared to the adult stage on both A. nerii and A. cyanophylli on potatoes when compared to larvae reared on these scales on squashes. Squash appears to be a less desirable and potato a more favourable host plant for survival and development. The results have implications for rearing programmes, and the release and establishment of C. nigritus, in fields and glasshouses where scale pests may be present on a variety of host plants, or may be on host plants different to those used in the insectary.

Multivariate survival analysis of histological parameters and clinical presentation in canine cutaneous mast cell tumours.

The Patnaik histological grade is a good method for the prediction of long-term mast cell tumour behaviour but it is influenced by subjective inter-observer variations and intratumoral heterogeneity. The present study evaluated each of the histopathological parameters used to formulate Patnaik's grade in terms of prognosis and tested whether they have a different prognostic sensitivity, thereby disclosing which could be considered more useful in the prediction of tumour recurrence and patient survival. Clinical presentation (single or multiple tumours) was also considered as possible prognostic factor. The results demonstrated that individual histological criteria together with multiple presentation may be of value in predicting the outcome of mast cell tumours. Among these, invasiveness (beta1.85; standard error 1.15) and the number of mitotic figures (beta3.01; standard error 1.18) showed high prognostic significance (Cox proportional hazard regression for censored data; chi-squared = 15.52, degree of freedom = 6, p = 0.016) and could serve as reliable prognostic indicators avoiding more subjective parameters such as cellular differentiation, nuclear morphology and tumoural pattern.

Acromegaly due to a somatroph adenoma in a dog.

A 10-year-old uncastrated male Dalmatian dog was referred for gait abnormalities consisting of chronic progressive stiffness and rigidity. Other symptoms were polyphagia associated with weight gain, polyuria and polydipsia, excessive panting, and an inspiratory stridor. The owner had noticed progressive thickening of the skin and enlargement of the tongue over the last 3 years. Physical examination revealed thickening of the skin, redundant skin folds, and enlargement of the tongue. The only remarkable abnormalities found on routine laboratory examination were mild anaemia and an increased serum fructosamine concentration. Circulating concentrations of total thyroxine, free thyroxine, and cTSH, and the results of an ACTH stimulation test were all within reference ranges. The basal serum growth hormone (GH) concentration was markedly elevated (23microg/l) and did not decrease during a glucose tolerance test or after somatostatin administration. The serum insulin-like growth factor-1 concentration was also markedly elevated (1254microg/l). Basal serum insulin concentration was high (95mU/l) and insulin concentrations increased considerably after glucose loading, consistent with insulin resistance. Abdominal ultrasonography showed no abnormalities. Survey radiographs of the vertebral column showed severe spondylosis deformans extending from the cervical to the lumbosacral spine. CT scanning of the skull showed an enlarged pituitary gland with normal enhancement pattern. On post-mortem examination, the entire vertebral column appeared as a single and inflexible structure due to the presence of multiple fused osteophytes. The pituitary gland contained an acidophilic adenoma that immunostained positively for GH (and negatively for ACTH and alpha-MSH). In conclusion, this Dalmatian dog with acromegaly and insulin resistance represents the first case of GH hypersecretion proven to be due to a somatotroph adenoma.

A comparison of methods to estimate cross-environment genetic correlations.

Advanced techniques for quantitative genetic parameter estimation may not always be necessary to answer broad genetic questions. However, simpler methods are often biased, and the extent of this determines their usefulness. In this study we compare family mean correlations to least squares and restricted error maximum likelihood (REML) variance component approaches to estimating cross-environment genetic correlations. We analysed empirical data from studies where both types of estimates were made, and from studies in our own laboratories. We found that the agreement between estimates was better when full-sib rather than half-sib estimates of cross-environment genetic correlations were used and when mean family size increased. We also note biases in REML estimation that may be especially important when testing to see if correlations differ from 0 or 1. We conclude that correlations calculated from family means can be used to test for the presence of genetic correlations across environments, which is sufficient for some research questions. Variance component approaches should be used when parameter estimation is the objective, or if the goal is anything other than determining broad patterns.

Spinal deformity in a sandtiger shark, Carcharias taurus Rafinesque: a clinical-pathological study.

The present study investigated spinal deformity in a captive sandtiger shark and describes the clinical and histopathological features of the lesion. Clinically, the shark presented a marked spinal curvature in the region between the pectoral girdle and cranial dorsal fin. Radiographic studies revealed scoliosis and kyphosis of the vertebral column with a modification of the basic structure of five vertebrae. Post-mortem examination confirmed the results of radiographic studies and histological examination showed remodelling and cartilaginous proliferation of the vertebral bodies together with necrosis, haemorrhage and signs of a local chronic inflammatory response extending to the surrounding muscle. Different aetiologies are proposed and discussed but nutritional imbalance (dietary vitamin deficiency) and/or traumatic injury are considered the most likely contributing factors.

E-cadherin and beta-catenin reduction influence invasion but not proliferation and survival in canine malignant mammary tumors.

E-Cadherin and beta-catenin are known for their role in tumor invasion, but both proteins also exert an influence on tumor proliferation. This study, performed on canine mammary tumors, aimed to analyze the influence of E-cadherin (E-cad) and beta-catenin (beta-cat), immunohistochemically assessed singly and in combination (E-cad/beta-cat), on survival and their relationship with several proliferation indices (AgNOR index, MIB1 index, mitotic index). Immunohistochemistry was carried out on 60 formalin-fixed, paraffin wax-embedded specimens of canine mammary malignancies. The labeling was defined as preserved when prevalent on cell membranes of more than 75% of cells and reduced in other forms of expression (i.e., membranous less than 75%, cytoplasmic, and negative). E-cad, beta-cat, and E-cad/beta-cat were preserved respectively in 22, 12, and 11 out of 60 cases. Immunohistochemical expression of the two proteins in the same tumors was significantly correlated (P = 0.0001; R = 0.57). Survival analysis revealed no difference in outcome comparing the preserved versus reduced cases (E-cad, P = 0.31; beta-cat, P = 0.29; E-cad/beta-cat P = 0.36). Grouping cases for histologic invasiveness, the expression of E-cad or beta-cat and E-cad/beta-cat showed a progressive reduction that paralleled an increase in invasiveness from noninfiltrating to stage-II tumors (E-cad, P < 0.001; beta-cat, P < 0.05; E-cad/beta-cat, P < 0.05). No significant difference was obtained comparing mitotic index, MIB 1 index, and AgNOR index by analysis of variance between the cases grouped for preserved or reduced E-cad, beta-cat, and E-cad/beta-cat variables. In conclusion, reduced expression of E-cad, beta-cat, or E-cad/beta-cat was significantly associated with the progression from noninfiltrating to highly infiltrating tumors but not with proliferation or survival.

Prognostic value of intratumoral vessel density in cutaneous mast cell tumors of the dog.

The biological behaviour of canine cutaneous mast cell tumours (MCTs) is highly variable. The aim of this study was to investigate whether intratumoral microvessel density (IMVD), as an indicator of angiogenesis, was predictive of biological behaviour in MCTs of canine skin, and to relate this parameter to histological grade and other histomorphological features. IMVD was evaluated by immunohistochemical labelling of the endothelial marker factor VIII (Von Willebrand's factor)-related antigen in 32 primary cutaneous MCTs. Computer image analysis was used to measure IMVD, expressed as the number of microvessels per square millimetre. Cases were assigned to low and high vessel density groups, the median value (14.1) being used as the "cut-off". Kaplan-Meier-estimated survival curves of the groups revealed that IMVD was significantly associated with survival time (P=0.046). One-way analysis of variance, performed to compare the IMVD values of tumours of different histological grades, showed a significant difference between grades I and III (P=0.0002). There was a significant difference in IMVD between moderately invasive and highly invasive tumours (P=0.002) and between tumours with different numbers of mitoses (P=0.0046). In multivariate analysis, performed to test the combined influence on survival of each parameter, only the mitotic index and IMVD were independent prognostic variables (P=0.00019). The relation of angiogenesis to biological behaviour and histological parameters of malignancy of cutaneous MCTs was consistent with previous findings in human and animal tumours.

Hypertrophy of intestinal smooth muscle in cats.

Pathological findings of four cats with severe and diffuse smooth muscle hypertrophy of the small intestine (MHSI) are reported and compared to those of five cats with segmental MHSI secondary to neoplastic obstruction and four controls. Histology demonstrated a constant association between idiopathic MHSI and submucosal fibrosis and chronic lymphoplasmacytic enteritis. Morphometry (gut diameter, thickness and area of muscular layers, number and density of smooth muscle nuclei) and MIB-1-immunolabelling showed that the thickness increase was mostly due to hypertrophy, but hyperplasia was also evident. Microbiology from ileal content samples was performed in two cats with primary MHSI, and Campylobacter spp. were isolated, which were also demonstrated by immunohistochemistry and ultrastructure. The association of chronic enteritis with idiopathic MHSI suggests that factors released in intestinal inflammation may also act as hypertrophy stimuli for smooth muscle cells.

E-cadherin expression in canine mammary carcinomas with regional lymph node metastases.

E-cadherin (E-cad) is a cell adhesion molecule known for its tumour invasion-suppressor function. This study investigated the immunohistochemical expression of E-cadherin in 19 cases of malignant mammary tumours of the dog and the relationship between E-cadherin expression in primary tumours and in regional lymph node metastases. E-cadherin expression is not always parallel in the primary tumour and in the lymph node metastasis. One year follow-up was available in 12 of 19 cases. Three different patterns of expression were revealed in the lymph node metastases compared with the primary tumour: downregulation when the protein expression was weaker in the metastasis than in the primary tumour; upregulation when E-cadherin was stronger in the lymph node than in the primary tumour, and a similarly intense expression when it was equal in the metastasis and in the tumour. The lymph node pattern revealed a prevalent upregulation or downregulation with respect to the primary tumour, whereas a similar expression of E-cadherin was encountered in less than 50% of cases.

Cell proliferation patterns in canine infundibular keratinizing acanthoma and well differentiated squamous cell carcinoma of the skin.

The aim of the present study was to investigate if the evaluation of cell proliferation of the well differentiated squamous cell carcinoma (WDSCC) and infundibular keratinizing acanthoma (IKA) could be useful in the differential diagnosis between these two tumours. Eighteen IKAs and ten WDSCCs were selected for this study. Two different methods were used to assess the activity of cell proliferation: MIB1 immunohistochemical detection and AgNOR proteins silver staining. The quantification of proliferative parameters was performed by means of an image analyzer and expressed as MIB1 index and AgNOR area (MNORA). Both MIBI immunohistochemical and AgNOR histochemical patterns were different in WDSCC and IKA; moreover analysis of variance showed a significant difference for both parameters employed (MIB1 index, MNORA) between WDSCC and IKA (P<0.003 for MIB1 index; P<0.0001 for AgNOR area). The results show that canine WDSCC and IKA have a different proliferative behaviour and the assessment of cell proliferation can be considered as a useful adjunctive tool to the histopathological investigation in the differential diagnosis of these tumours.

Quantitative genetics of signal evolution: a comparison of the pheromonal signal in two populations of the cabbage looper, Trichoplusia ni.

Pheromones are important in reproductive isolation among populations of moths, but the genetics associated with diversification of pheromonal signals is poorly understood. To gain insight into processes that may lead to diversification we examined the genetic architecture underlying the production of the sex pheromone of the cabbage looper moth, Trichoplusia ni. We compared genetic parameters of two populations; one with a wild-type pheromone phenotype (N) and one where a single-gene mutation affecting the pheromone blend produced by females had been established (M). Using a half-sib breeding design we estimated heritabilities, coefficients of additive genetic variation, and phenotypic, genetic, and environmental correlations of the pheromone components. In both populations, narrow sense heritabilities were generally moderate and genetic correlations were mostly positive. Comparisons between the two populations showed that, while the pattern of phenotypic correlations showed significant agreement between populations, the patterns of genetic (co)variation (i.e. the shapes of the within population matrix) were dissimilar between the two populations. The presence of additive genetic variation in both populations indicates that there is the potential for further evolution of individual pheromone components. However, because of the differences between the populations in the pattern of genetic variation and covariation, the populations will evolve along different evolutionary trajectories even under identical selection pressures. These results suggest that single gene mutations, once established, can be associated with further alterations in the genetic architecture and this has implications for the evolution of pheromone communication.

Rapid methods to extract DNA and RNA from Cryptococcus neoformans.

Extraction of nucleic acids from the pathogenic yeast Cryptococcus neoformans is normally hampered by a thick and resistant capsule, accounting for at least 70% of the whole cellular volume. This paper presents procedures based on mechanical cell breakage to extract DNA and RNA from C. neoformans and other capsulated species. The proposed system for DNA extraction involves capsule relaxation by means of a short urea treatment and bead beating. These two steps allow a consistent extraction even from strains resistant to other procedures. Yield and quality of DNA obtained with the proposed method were higher than those obtained with two earlier described methods. This protocol can be extended to every yeast species and particularly to those difficult to handle for the presence of a capsule. RNA purification is accomplished using an original lysing matrix and the FastPrep System (Bio101) after a preliminary bead beating treatment. Yields range around 1 mg RNA from 15 ml overnight culture (10(9) cells), RNA appears undegraded, making it suitable for molecular manipulations.