Structural and functional characterization of myotoxin I, a Lys49 phospholipase A(2) homologue from Bothrops moojeni (Caissaca) snake venom

Myotoxin-I (MjTX-I) was purified to homogeneity from the venom of Bothrops moojeni by ion-exchange chromatography on CM-Sepharose. Its molecular weight, estimated by SDS-PAGE, was 13,400 (reduced) or 26,000 (unreduced). The extinction coefficient (E-1.0 cm(1.0 mg/ml)) of MjTX-I was 1.145 at lambda = 278 nm, pH 7.0, and its isoelectric point was 8.2 at ionic strength mu = 0.1. When lyophilized and stored at 4 degrees C, dimeric, trimeric, and pentameric forms of the protein were identified by SDS-PAGE. This heterogeneous sample could be separated into three fractions by gel filtration on Sephadex 6-50. The fractions were analyzed by isoelectric focusing, immunoelectrophoresis, and amino acid composition, which indicated that heterogeneity was the result of different levels of self-association. Protein sequencing indicated that MjTX-I is a Lys49 myotoxin and consists of 121 amino acids (M-r = 13,669), containing a high proportion of basic and hydrophobic residues. It shares a high degree of sequence identity with other Lys49 PLA(2)-like myotoxins, but shows a significantly lower identity with catalytically active Asp49 PLA(2)s. The three-dimensional structure of MjTX-I was modeled based on the crystal structures of three highly homologous Lys49 PLA(2)-like myotoxins. This model showed that the amino acid substitutions are conservative, and mainly the beta-wing region, and the C-terminal extended random coil. MjTX-I displays local myotoxic and edema-inducing activities in mice, and is lethal by intraperitoneal injection, with an LD50 value of 8.5 +/- 0.8 mg/kg, In addition, it is cytotoxic to myoblasts/ myotubes in culture, and disrupts negatively charged liposomes. In comparison with the freshly prepared dimeric sample, the more aggregated forms showed significantly reduced myotoxic activity. However, the edema-inducing activity of MjTX-I was independent of molecular association. Phospholipase A(2) activity on egg yolk, as well as anticoagulant activity, were undetectable both in the native and in the more associated forms. His, Tyr, and Trp residues of the toxin were chemically modified by specific reagents. Although the myotoxic and lethal activities of the modified toxins were reduced by these treatments, neither its edema-inducing or Liposome-disrupting activities were significantly altered. Rabbit antibodies to native MjTX-I cross-reacted with the chemically modified forms, and both the native and modified MjTX-I preparations were recognized by antibodies against the C-terminal region 115-129 of myotoxin II from B. asper, a highly Lys49 PLA(2)-homologue with high sequencial similarity. (C) 2000 Academic Press.

Morphology of the nymphoid replacement reproductives in the neotropical termite Armitermes euamignathus (Isoptera, Termitidae, Nasutitermitinae)

Numerous nymphoid reproductives were found in three field nests of Armitermes euamignathus collected in Brazil. We report here a morphological description and a biometric study of these individuals. Nymphoid replacements displayed narrow wing buds when compared with those present in nymphs from the three last instars. Thorax morphology of the nymphoids was similar to the penultimate nymphal instar (N4) or to the ultimate nymphal instar (N5), and their origin from these instars of nymphs is discussed. All the nymphoids had eyes, ocelli, and 15 antennal segments. The nymphoid females from nest 1 had different grades of physogastry and royal fat body. The nymphoid females from nests 2 and 3, the nymphoid males from all nests, and the primary king from nests 2 acid 3 had a common fat body, which is similar to that present in alates. The ovaries and the testes of nymphoids were fully mature and the corpora allata larger than those in imagoes. The mandibular glands were also enlarged in nymphoids but the tergal glands were absent. (C) 1999 Wiley-Liss, Inc.

Induction of the split sting trait in Africanized Apis mellifera (Hymenoptera : Apidae) by cold treatment of pupae

Split sting is the name given to a nonfunctional honey bee sting characterized by lancets not attached to the stylet. It has appeared in a mutant line in Brazil, and has provoked interest as a possible means to reduce honey bee colony defensiveness. We induced this alteration in Africanized Apis mellifera L. workers and queens by maintaining pupae at 20 degrees C. In particular, we determined the pupal phase most susceptible to alterations in the sting caused by cold treatment, and we investigated whether this treatment also affected survival to the adult phase and wing morphology. The highest frequency of split sting was detected in workers treated at the pink-eyed pupal phase. The lowest frequency was observed in the bees treated at the oldest worker pupal phase studied (brown-eyed pupae with lightly pigmented cuticle). Both queen pupal phases tested (white and pink-eyed pupae) were equally sensitive and produced high percentages of adults with split sting. However, the 20 degrees C treatment of workers and queens, at the different pupal phases, resulted in high frequencies of adults with deformed wings. Also, fewer workers and queens treated at the earlier pupal stages reached adult emergence. There was also an arrest in developmental time, corresponding to the period of cold treatment.

Assessment of brachial plexus blockade in chickens by an axillary approach

Objective To assess the brachial plexus block in chickens by an axillary approach and using a peripheral nerve stimulator.Study design Prospective, randomized, double-blinded study.Animals Six, 84-week old, female chickens.Methods Midazolam (1 mg kg(-1)) and butorphanol (1 mg kg(-1)) were administered into the pectoralis muscle. Fifteen minutes later, the birds were positioned in lateral recumbency and following palpation of the anatomic landmarks, a catheter was inserted using an axillary approach to the brachial plexus. Lidocaine or bupivacaine (1 mL kg(-1)) was injected after plexus localization by the nerve stimulator. Sensory function was tested before and after blockade (carpus, radius/ulna, humerus and pectoralis muscle) in the blocked and unblocked wings. The latency to onset of motor and sensory block and the duration of sensory block were recorded. A Friedman nonparametric one-way repeated-measures ANOVA was used to compare scores from baseline values over time and to compare the differences between wings at each time point.Results A total of 18 blocks were performed with a success rate of 66.6% (12/18). The latency for motor block was 2.8 +/- 1.1 and 3.2 +/- 0.4 minutes for lidocaine and bupivacaine, respectively. The latencies for and durations of the sensory block were 6.0 +/- 2.5 and 64.0 +/- 18.0 and 7.8 +/- 5.8 and 91.6 +/- 61.7 minutes for lidocaine and bupivacaine, respectively. There was no statistical difference between these times for lidocaine or bupivacaine. Sensory function was not abolished in nonblocked wings.Conclusions and clinical relevance The brachial plexus block was an easy technique to perform but had a high failure rate. It might be useful for providing anesthesia or postoperative analgesia of the wing in chickens and exotic avian species that have similar wing anatomy.

ARE CORPORAL ALLOMETRIC MEASUREMENTS GOOD PREDICTORS of PALATABILITY IN NEOTROPICAL BUTTERFLIES (LEPIDOPTERA)?

Since palatable butterflies are more dependent on evasive flight to escape from predators, they should be more restricted in their flight-related morphology than unpalatable ones. We compared: the ratios between the (1) length of head plus thorax and the length of abdomen (A/B), (2) length of the tip of the head to wing base and the length of the wing base to end of the abdomen (C/D), (3) the variances of A/B and C/D, (4) the proportion between the thoracic and the body weight, and (5) the flight speed between palatable and unpalatable butterflies. A/B and thoracic/body weight were higher for palatable species, indicating higher body symmetry and muscular mass. However, flight speed did not differ. Unexpectedly, the variance of A/B was higher for palatable species while that of C/D did not differ. Therefore, corporal allometric measurements of Neotropical butterflies are good predictors of palatability, though not of flight speed.

Predicting body and carcass characteristics of 2 broiler chicken strains using support vector regression and neural network models

As a new modeling method, support vector regression (SVR) has been regarded as the state-of-the-art technique for regression and approximation. In this study, the SVR models had been introduced and developed to predict body and carcass-related characteristics of 2 strains of broiler chicken. To evaluate the prediction ability of SVR models, we compared their performance with that of neural network (NN) models. Evaluation of the prediction accuracy of models was based on the R-2, MS error, and bias. The variables of interest as model output were BW, empty BW, carcass, breast, drumstick, thigh, and wing weight in 2 strains of Ross and Cobb chickens based on intake dietary nutrients, including ME (kcal/bird per week), CP, TSAA, and Lys, all as grams per bird per week. A data set composed of 64 measurements taken from each strain were used for this analysis, where 44 data lines were used for model training, whereas the remaining 20 lines were used to test the created models. The results of this study revealed that it is possible to satisfactorily estimate the BW and carcass parts of the broiler chickens via their dietary nutrient intake. Through statistical criteria used to evaluate the performance of the SVR and NN models, the overall results demonstrate that the discussed models can be effective for accurate prediction of the body and carcass-related characteristics investigated here. However, the SVR method achieved better accuracy and generalization than the NN method. This indicates that the new data mining technique (SVR model) can be used as an alternative modeling tool for NN models. However, further reevaluation of this algorithm in the future is suggested.

Experimental Study on the Impact Behaviour of Metal-fiber Laminates

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Geographic variation of Africanized honey bees (Apis mellifera L.) in Brazil: Multivariate morphometrics and racial admixture

The correspondence between morphometric and isozymic geographic variation patterns of Africanized honey bees in Brazil was analyzed. Morphometric data consisted of mean vectors of 19 wing traits measured in 42 local populations distributed throughout the country. Isozymic data refer to allelic frequencies of malate dehydrogenase (MDH), and were obtained from Lobo and Krieger. The two data sets were analyzed through canonical trend surface, principal components and spatial autocorrelation analyses, and showed north-south dines, demonstrating that Africanized honey bees in southern and southeastern Brazil are more similar to European honey bees than those found in northern and northeastern regions. Also, the morphometric variation is within the limits established by the racial admixture model, considering the expected values of Africanized honey bee fore wing length (WL) in southern and northeastern regions of Brazil, estimated by combining average values of WL in the three main subspecies involved in the Africanization process (Apis mellifera scutellata, A. m. ligustica and A. m. mellifera) with racial admixture coefficients.

Efeitos de diferentes densidades populacionais nas estações fria e quente do ano sobre o desempenho de frangos de corte

The objective of this study was to evaluate the performance of broilers reared under different population densities in the cold and hot seasons of the year. Two identical experiments were conducted, one during the winter for 49 days, and the other during the summer where the chicks were slaughtered at 42 days of age. Commercial Hubbard broiler-type chicks were distributed in a randomized block design in a 4 × 2 factorial arrangement with population densities of 10, 14, 18, and 22 birds/m 2, by sex, and with four replications. Feed intake was reduced with a progressive decrease of available space for the chicks, and it resulted in a linear decrease in weight gain. However, there was a linear increase in the live weight of broilers in kilograms per area of floor space, proportional to the increase of population density, without effect on the viability. The progressive increment in the population density reduced the observed feed/gain ratio during the total winter rearing period for both sexes. There was no effect of population density on this characteristic during the hot season of the year. Population density did not affect the carcass yield of chicks during the summer. However, there was a linear increase on carcass yield with an increase in population density in the winter. The males, at the age slaughtered, had higher weights, feed intakes, and better feed/gain ratios, and lower abdominal fat and higher live weight production in kilograms per floor area than females. However, the viable index for the males was lower. From the results obtained, there was a linear increase in the liveweight of broilers in kilograms per floor area, making it possible to rear broilers under higher population densities, independent of the season of the year, as well as the occurrence of a decrease in feed intake and weight gain of the chicks with a decrease of available space.

Response to orphaning in two Neotropical termites: Armitermes euamignathus and Embiratermes festivellus

In this paper we examine the potential of the termites Armitermes euamignathus Silvestri: 1901 and Embiratermes festivellus (Silvestri, 1901) (Isoptera, Termitidae, Nasutitermitinae) to produce neotenics experimentally. Three nests of the mound-building termite A. euamignathus, from the Brazilian cerrado, had their primary queens removed in August 1994. After 12 months, only one mound survived; it had a normal appearance. In this healthy, orphaned colony we found the primary king, six physogastric nymphoid female replacement reproductives, two ergatoid female replacement reproductives, 46 nymphs, several presoldiers, soldiers, workers, larvae and many eggs. These data show that neotenics in A. euamignathus may originate from both workers and nymphs, but nymphoids are produced in larger numbers. The biometric study of nymphs and nymphoids suggests that these brachypterous neotenics were derived from third instar nymphs after a single moult or from four instar nymphs after a reduction of wing bud length. A piece of an E. festivellus nest with some third instar nymphs, soldiers and workers was kept under laboratory conditions. After 12 months, the whole experimental subcolony was examined and appeared to contain two pigmented nymphoid females, two pigmented nymphoid males, only one larva, seven nymphs of the same instar, 148 workers, five soldiers and many eggs. These results also indicate the capacity of the termite E. festivellus to produce nymphoid neotenics. These neotenic females were laying eggs, but they were not physogastric after a year, unlike some nymphoids of the same species collected from natural colonies.

Morphometric variations in the housefly, Musca domestica (L.) with latitude

Morphologic analysis of geographic strains of Musca domestica carried out on natural and laboratory experiments starting with 400 and 800 eggs showed phenotypic variations related with latitude. Females of the natural populations showed clines for several morphological traits of the wing, whereas male flies showed a reduction in the dispersion measures (s2 and CV) of wing width and length. The same reduction was obtained for males, females and total number of flies of the natural populations in dispersion of the number of bristles on the fourth abdominal sternite. A significant negative correlation was observed for the head width of females and for the total number of flies emerged in the laboratory experiments started with 400 eggs. All flies produced by the experiments starting with 800 eggs showed a reduction in variability of dispersion of the bristles on the fourth abdominal sternite in the strains obtained from locations south of the area analyzed. Evolutionary aspects of these correlation coefficients between morphometric traits and latitude are discussed.

Affinities of the Saw-billed Hermit (Ramphodon naevius) determined by cytochrome-b sequence data

We sequenced 912 bp of the cytochrome-b gene to examine phylogenetic relationships of the enigmatic Saw-billed Hermit (Ramphodon naevius), a large and distinctive hummingbird endemic to tropical forests of southeastern Brazil. Bootstrapped maximum parsimony and maximum likelihood analyses of sequence data from 11 hummingbirds and several outgroups (two swifts, one goatsucker) support: (a) monophyly of the traditional hermit (Phaethornithinae) and nonhermit (Trochilinae) subfamilies, (b) placement of Ramphodon among hermits, and (c) a sister relationship between Ramphodon and an exemplar of the widespread polytypic hermit genus Glaucis. The association of Ramphodon with derived hermit lineages is concordant with subfamilial patterns of wing anatomy and nest architecture. However, the unusual plumages (striped underparts) and male bills (long, serrated, hooked) shared by Ramphodon and the Tooth-billed Hummingbird (Androdon aequatorialis) appear to have evolved within separate hermit and nonhermit tooth-billed clades. Distal placement of the Ramphodon-Glaucis clade within hermits implies that even distinctive Brazilian endemics such as Ramphodon are derived forms that evolved relatively recently.

Comparative study of the cuticular hydrocarbon composition of Melipona bicolor Lepeletier, 1836 (Hymenoptera, Meliponini) workers and queens

In social insects, cuticular hydrocarbons are involved in species, kin, caste and nestmate recognition. Gas chromatography and mass spectrometry were used to compare the cuticular hydrocarbon composition of workers, males and queens of Melipona bicolor. The cuticular hydrocarbon composition of this species was found to consist mainly of C23, C25:1, C25, C27:1, C27, C29:1 and C29, which are already present in imagoes that have not yet abandoned the brood cell. This composition varied quantitatively and qualitatively between and within the castes and sexes. The newly emerged workers and young queens (virgins) had similar cuticular hydrocarbon profiles, which were different from those of the males. When the females start executing their tasks in the colony, the cuticular hydrocarbon profile differences appear. The workers have less variety, while the queens conserve or increase the number of cuticular hydrocarbon compounds. The queens have more abdominal tegumentary glands than the workers, which apparently are the source of the new cuticular compounds.

Microdon tigrinus (Diptera, Syrphidae): Populational fluctuations and specificity to the nest of Acromyrmex coronatus (Hymenoptera: Formicidae)

This study was performed in the southern and southeastern regions of Brazil, visiting numerous municipalities, over a period of 25 years, excavating nests of 12 species of leaf-cutting ants of the Acromyrmex genus. Larvae and pupae of Microdon tigrinus were found only in nests of the leaf-cutting ant Acromyrmex coronatus, indicating high specificity. Observation showed that larvae and pupae were well accepted in the nests and the adults, immediately after puparia eclosion and prior to wing distension, were not attacked by the workers, suggesting that they produce semiochemicals for a short time period until they arrive outside the Acromyrmex coronatus nest. It was postulated that these larvae feed on the organic detritus of the nest, as shown for Microdon larvae of other species.

Osteologia craniana de Nyctibiidae (Aves, Caprimulgiformes)

The Potoos form an exclusively neotropical family of nocturnal birds distributed throughout Central and South America, except Chile, and reaching their highest diversity in the Amazon region. The seven currently recognized species are certainly among the most poorly known birds of this region. They are characterized by a distinctive mimicry of vegetal trunks, where they remain almost motionless during daytime. For this reason, their nocturnal and cryptic habits make them exceedingly difficult to study. Published accounts on behavior and natural history of the family are scarce and contributions regarding its anatomy are rare. Here we sample six of the seven currently recognized species of Nyctibiidae, including Nyctibius grandis, N. aethereus, N. griseus, N. jamaicensis, N. leucopterus and N. bracteatus, in order to conduct a detailed and illustrated description of the skull and jaw osteology. High interspecific variation in skull osteology was observed in the family. Species of this family possess a highly modified skull, adapted to their life habits, which shelters their well developed eyes and permits a large mouth opening. The bones that form the palate structure exhibit a dorsoventral flattening, particularly in the pterigoid and parasphenoid bones, with the palatine bone being a broadly developed, wing-shaped structure. In the maxilar region, near the jugal arch, there is a tooth-like projection, unique among birds, which may assist in the retention of prey upon capture. The vomer bone is highly variable within the family, showing varying numbers of rostral projections amongst species. The broad occipital region exhibits large spacing between the quadrate bones, which are vertically disposed and possess a reduced processus orbitalis. The mandible, which is flexible and elastic, has an extremely short symphyseal region and sindesmotic joints in both mandibular rami. As a family, potoos possess a highly specialized skull which provides insight into the relationship between the form of the structures and the feeding habits of the species. Furthermore, the large interspecific variation in skull morphology emphasizes the needs for taxonomic revision within the family, which at present is lumped into a single genus.