LOCALIZATION OF DIPLOPTERA-PUNCTATA ALLATOSTATIN-LIKE IMMUNOREACTIVITY IN HELMINTHS - AN IMMUNOCYTOCHEMICAL STUDY

The nervous systems of helminths are predominantly peptidergic in nature, although it is likely that the full range of regulatory peptides used by these organisms has yet to be elucidated. Attempts to identify novel helminth neuropeptides are being made using immunocytochemistry with antisera raised against peptides isolated originally from insects. One of these antisera was raised against allatostatin III, a peptide isolated originally from the cockroach, Diploptera punctata, and a member of a family of related peptides found in insects. Allatostatin immunoreactivity was found throughout the nervous systems of Mesocestoides corti tetrathyridia, and adult Moniezia expansa, Diclidophora merlangi, Fasciola hepatica, Schistosoma mansoni, Ascaris suum and Panagrellus redivivus. Immunostaining was observed in the nerve cords and anterior ganglia of all the helminths. It was also apparent in the subtegumental nerves and around the reproductive apparatus of the flatworms, in neurones in the pharynx of D. merlangi, F. hepatica, A. suum and P. redivivus, and in fibres innervating the anterior sense organs in the nematodes. Immunostaining in all species was both reproducible and specific in that it could be abolished by pre-absorption of the antiserum with allatostatins I-IV. These results suggest that molecules related to the D. punctata allatostatins are important components in the nervous systems of a number of helminth parasites, and a free-living nematode. Their distribution within the nervous system suggests they function as neurotransmitters/ neuromodulators with roles in locomotion, feeding, reproduction and sensory perception.

PEPTIDES RELATED TO THE DIPLOPTERA-PUNCTATA ALLATOSTATINS IN NONARTHROPOD INVERTEBRATES - AN IMMUNOCYTOCHEMICAL SURVEY

The allatostatins are a family of peptides isolated originally from the cockroach, Diploptera punctata. Related peptides have been identified in Periplaneta americana and the blowfly, Calliphora vomitoria. These peptides have been shown to be potent inhibitors of juvenile hormone synthesis in these species. A peptide inhibitor of juvenile hormone biosynthesis has also been isolated from the moth, Manduca sexta; however, this peptide has no structural homology with the D. punctata-type allatostatins. Investigations of the phylogeny of the D. punctata allatostatin peptide family have been started by examining a number of nonarthropod invertebrates for the presence of allatostatin-like molecules using immunocytochemistry with antisera directed against the conserved C-terminal region of this family. Allatostatin-like immunoreactivity (ALIR) was demonstrated in the nervous systems of Hydra oligactis (Hydrozoa), Moniezia expansa (Cestoda), Schistosoma mansoni (Trematoda), Artioposthia triangulata (Turbellaria), Ascaris suum (Nematoda), Lumbricus terrestris (Oligochaeta), Limax pseudoflavus (Gastropoda), and Eledone cirrhosa (Cephalopoda). ALIR could not be demonstrated in Ciona intestinalis (Ascidiacea). These results suggest that molecules related to the allatostatins may play an important role in nervous system function in many invertebrates as well as in insects and that they also have an ancient evolutionary lineage. (C) 1994 Wiley-Liss, Inc.

Molting dynamics and juvenile hormone titer profiles in the nymphal stages of a lower termite, Cryptotermes secundus (Kalotermitidae) - signatures of developmental plasticity

Termites are social cockroaches and this sociality is founded on a high plasticity during development. Three molting types (progressive, stationary and regressive molts) are fundamental to achieve plasticity during alate/sexual development, and they make termites a major challenge to any model on endocrine regulation in insect development. As the endocrine signatures underpinning this plasticity are barely understood, we studied the developmental dynamics and their underlying juvenile hormone OH) titers in a wood-dwelling termite. Cryptotermes secundus, which is characterized by an ancestral life style of living in dead wood and individuals being totipotent in development. The following general pattern elements could be identified during winged sexual development (i) regressive molts were accompanied by longer intermolt periods than other molting types, (ii) JH titers decreased gradually during the developmental transition from larva (immatures without wing buds), to nymph (immatures with wing buds), to winged adult, (iii) in all nymphal stages, the JH titer rose before the next molt and dropped thereafter within the first week, (iv) considerable variation in JH titers occurred in the midphase of the molting cycle of the 2nd and 3rd nymphal instar, inferring that this variation may reflect the underlying endocrine signature of each of the three molting types, (v) the 4th nymphal instar, the shortest of all, seems to be a switch point in development, as nymphs in this stage mainly developed progressively. When comparing these patterns with endocrine signatures seen in cockroaches, the developmental program of Cryprotermes can be interpreted as a co-option and repetitive use of hormonal dynamics of the post dorsal-closure phase of cockroach embryonic development. (C) 2012 Elsevier Ltd. All tights reserved.

A cytochrome P450 terpenoid hydroxylase linked to the suppression of insect juvenile hormone synthesis

A cDNA encoding a cytochrome P450 enzyme was isolated from a cDNA library of the corpora allata (CA) from reproductively active Diploptera punctata cockroaches. This P450 from the endocrine glands that produce the insect juvenile hormone (JH) is most closely related to P450 proteins of family 4 and was named CYP4C7. The CYP4C7 gene is expressed selectively in the CA; its message could not be detected in the fat body, corpora cardiaca, or brain, but trace levels of expression were found in the midgut and caeca. The levels of CYP4C7 mRNA in the CA, measured by ribonuclease protection assays, were linked to the activity cycle of the glands. In adult females, CYP4C7 expression increased immediately after the peak of JH synthesis, reaching a maximum on day 7, just before oviposition. mRNA levels then declined after oviposition and during pregnancy. The CYP4C7 protein was produced in Escherichia coli as a C-terminal His-tagged recombinant protein. In a reconstituted system with insect NADPH cytochrome P450 reductase, cytochrome b5, and NADPH, the purified CYP4C7 metabolized (2E,6E)-farnesol to a more polar product that was identified by GC-MS and by NMR as (10E)-12-hydroxyfarnesol. CYP4C7 converted JH III to 12-trans-hydroxy JH III and metabolized other JH-like sesquiterpenoids as well. This ω-hydroxylation of sesquiterpenoids appears to be a metabolic pathway in the corpora allata that may play a role in the suppression of JH biosynthesis at the end of the gonotrophic cycle.

Seasonal growth of King George whiting (Sillaginodes punctata) estimated from length-at-age samples of the legal-size harvest

Samples of 11,000 King George whiting (Sillaginodes punctata) from the South Australian commercial and recreational catch, supplemented by research samples, were aged from otoliths. Samples were analyzed from three coastal regions and by sex. Most sampling was undertaken at fish processing plants, from which only fish longer than the legal minimum length were obtained. A left-truncated normal distribution of lengths at monthly age was therefore employed as model likelihood. Mean length-at-monthly-age was described by a generalized von Bertalanffy formula with sinusoidal seasonality. Likelihood standard deviation was modeled to vary allometrically with mean length. A range of related formulas (with 6 to 8 parameters) for seasonal mean length at age were compared. In addition to likelihood ratio tests of relative fit, model selection criteria were a minimum occurrence of high uncertainties (>20% SE), of high correlations (>0.9, >0.95, and >0.99) and of parameter estimates at their biological limits, and we sought a model with a minimum number of parameters. A generalized von Bertalanffy formula with t0 fixed at 0 was chosen. The truncated likelihood alleviated the overestimation bias of mean length at age that would otherwise accrue from catch samples being restricted to legal sizes.

Biology and morphometry study of Drepane punctata (Cuv and Val) at juvenile stage of Bombay coast

The metric and meristic characters of Drepane punctata from Versova Fish Landing Centre of Bombay have been studied. The meristic characters showed considerable variations. The number of dorsal fin rays were relatively stable characters. Statistical interpretation of metric data indicated that there is a direct relationship between the preanal, predorsal in relation to total length and eye-diameter in relation to head length.

Length-weight relationship of Drepane punctata (Cuv. & Val.) at juvenile stage off Bombay coast

The study of the length weight relationship is highly useful to the fishery biologists in the study of population dynamics of fish and for determining the pattern of growth of stock. The parabolic equation for the length-weight relationship of Drepane punctata juveniles off the Bombay coast, expresses the value of "n" 2.83, indicating that the growth rate is less than the cube length.

Aspectos biológicos de Orphulella punctata (De Geer) (Orthoptera:Acrididae)

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Efeitos de herbicidas nos teores de clorofilas de Spirodela punctata

Com o objetivo de verificar na planta aquática Spirodela punctata o efeito de diferentes concentrações (0,005; 0,05; 0,5 e 5 mgL-1 de ingrediente ativo) dos herbicidas butachlor, glyphosate e propanil sobre os teores de clorofila a, b e na razão clorofila a/b, foram conduzidos, por sete dias completos, bioensaios em sala climatizada. Determinou-se os teores de clorofilas através de leituras espectrofotométricas nas absorbâncias A645 e A663 nm. Os resultados mostraram que o butachlor diminuiu o teor de clorofila b, o glyphosate o teor de clorofila a, enquanto o propanil diminuiu os teores de clorofila a, b e a razão a/b, provocando a maior redução de pigmentos na espécie.

The extract of the jellyfish Phyllorhiza punctata promotes neurotoxic effects

Phyllorhiza punctata (P. punctata) is a jellyfish native to the southwestern Pacific. Herewith we present the biochemical and pharmacological characterization of an extract of the tentacles of P. punctata. The tentacles were subjected to three freezethaw cycles, homogenized, ultrafiltered, precipitated, centrifuged and lyophilized to obtain a crude extract (PHY-N). Paralytic shellfish poisoning compounds such as saxitoxin, gonyautoxin-4, tetrodotoxin and brevetoxin-2, as well as several secretory phospholipase A2 were identified. PHY-N was tested on autonomic and somatic neuromuscular preparations. In mouse vas deferens, PHY-N induced phasic contractions that reached a peak of 234 +/- 34.7% of control twitch height, which were blocked with either 100 mu m of phentolamine or 1m m of lidocaine. In mouse corpora cavernosa, PHY-N evoked a relaxation response, which was blocked with either L-NG-Nitroarginine methyl ester (0.5 m m) or 1m m of lidocaine. PHY-N (1, 3 and 10 mu g ml(-1)) induced an increase in tonus of the biventercervicis neuromuscular preparation that was blocked with pre-treatment of galamine (10 mu m). Administration of 6 mg kg(-1) PHY-N intramuscularly produced death in broilers by spastic paralysis. In conclusion, PHY-N induces nerve depolarization and nonspecifically increases neurotransmitter release. Copyright (C) 2011 John Wiley & Sons, Ltd.

Immunological responses and cytokine gene expression analysis to Cooperia punctata infections in resistant and susceptible Nelore cattle

Cellular and humoral immune response, as well as cytokine gene expression, was assessed in Nelore cattle with different degrees of resistance to Cooperia punctata natural infection. One hundred cattle (male, weaned, 11-12 months old), kept together on pasture, were evaluated. Faecal and blood samples were collected for parasitological and immunological assays. Based on nematode faecal egg counts (FEC) and worm burden, the seven most resistant and the eight most susceptible animals were selected. Tissue samples of the small intestine were collected for histological quantification of inflammatory cells and analysis of cytokine gene expression (IL-2, IL-4, IL-8, IL-1 2p35, IL-13, TNF-alpha, IFN-gamma, MCP-1, MCP-2, and MUC- 1) using real-time RT-PCR. Mucus samples were also collected for IgA levels determination. Serum IgG1 mean levels against C. punctata antigens were higher in the resistant group, but significant differences between groups were only observed 14 days after the beginning of the experiment against infective larvae (1-3) and 14 and 84 days against adult antigens. The resistant group also presented higher IgA levels against C. punctata (L3 and adult) antigens with significant difference 14 days after the beginning of the trial (P < 0.05). In the small-intestine mucosa, levels of IgA anti-L3 and anti-adult C. punctata were higher in the resistant group, compared with the susceptible group (P < 0.05). Gene expression of both T(H)2 cytokines (IL-4 and IL-13) in the resistant group and T(H)1 cytokines (IL-2, IL-1 2p35, IFN-gamma and MCP-1) in the susceptible group was up-regulated. Such results suggested that immune response to C. punctata was probably mediated by TH2 cytokines in the resistant group and by T(H)1 cytokines in the susceptible group. (C) 2008 Elsevier B.V. All rights reserved.

Ensaios sobre a metodologia do estudo da influência de três herbicidas em Spirodela punctata (G.F.W. Meyer) Thompson (Lemnaceae)

O presente trabalho visa conhecer a taxa de multiplicação e o índice de mortalidade de S. punctata a diversas concentrações do butaclora, propanila e glifosato bem como comparar dois métodos diferentes de investigações (ensaio 1 e 2). Os dados indicaram que no ensaio 1 as concentrações foram pouco tóxicas provocando baixos índices de mortalidade embora tenha ocorrido inibição do crescimento nas concentrações mais fortes. O ensaio 2 mostrou que a butaclora e a propanila são mais tóxicas que o glifosato, com valores de mortalidade de 100% em apenas 48 horas. A taxa de multiplicação, neste ensaio, somente foi determinada para o glifosato, pois os demais herbicidas foram altamente tóxicos para a espécie. Verificou-se, na comparação dos dois métodos diferentes, que os recipientes utilizados no ensaio 2 foram mais adequados ao cultivo da espécie e que os efeitos tóxicos devem ser observados em frondes consideradas individualmente e não em plantas inteiras.

Distribution and habitat use of Sordellina punctata (Serpentes, Colubridae), with a new record from State of São Paulo, Brazil

Sordellina punctata occurs throughout south and southeastern Brazil in the Atlantic Forest domain (states of São Paulo, Paraná and Santa Catarina). Records from the states of Rio de Janeiro and Mato Grosso do Sul are questionable. A new record from southwestern São Paulo extends its distribution west and northward, probably indicating a wider range in the past. Habitat use in S. punctata implies that it may be more associated with wetlands and other saturated areas than truly aquatics ones.

Resistência de Haemonchus placei, Cooperia punctata e Oesophagostomum radiatum à ivermectina pour-on a 500mcgkg-1 em rebanhos bovinos no Brasil

The broad spectrum of endectocides and the easy use mode of their pour-on formulations are factors that have stimulated a higher frequency of use these drugs in cattle. In this study was evaluated the efficacy of ivermectin pour-on using the dose of 500mcgkg(-1), against nematodes in naturally infected cattle from different herds. Twelve calves were brought from each of the four farms selected. All the 48 calves used showed mean of eggs per gram of feces (EPG) greater than 500 considering three consecutive counts. Subsequently, animals from each herd were divided into two groups of six animals each, one treated pour-on with ivermectin 500mcgkg(-1) and other kept as untreated control. Calves were euthanized 14 days after treatment for counting the endoparasites. Ivermectin showed null effect against H. placei in all the herds evaluated. The drug was also ineffective against C. punctata in the herds from Jaboticabal, SP and Formiga, MG, and reached efficacy of 75.8% and 58.4% in herds from Sao Jose do Rio Pardo, SP and Sao Sebastiao do Para so, MG, respectively. Efficacies of 94.2% (Jaboticabal), 0.0% (Sao Jose do Rio Pardo), 94.2% (Formiga) and 39.2% (Sao Sebastiao do Para so) were detectedagainstO. radiatum. Based on these results obtained on the present study, the four populations of Haemonchus placei and Cooperia punctata evaluated were resistant to ivermectin pour-on using a dose of 500mcgkg(-1). Ivermectin-resistant strains of Oesophagostomum radiatum were found in two of the four herds evaluated.

Enriquecimento de amido de biomassa produzida em lagoa de lemna utilizando a espécie Landoltia punctata

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