Survival, population pize, and gonotrophic cycle duration of Nyssomyia neivai (Diptera: Psychodidae) at an endemic area of American Cutaneous Leishmaniasis in Southeastern Brazil

The survival, absolute population size, gonotrophic cycle duration, and temporal and spatial abundance of Nyssomyia neivai (Pinto) were studied in a rural area endemic for American cutaneous leishmaniasis (ACL) in Conchal, Sõo Paulo State, southeastern Brazil, using mark-release-recapture techniques and by monitoring population fluctuation. The monthly abundance exhibited a unimodal pattern, with forest and domicile habitats having the highest relative abundances. A total of 1,873 males and 3,557 females were marked and released during the six experiments, of which 4.1-13.0 per cent of males and 4.1-11.8 per cent of females were recaptured. Daily survivorship estimated from the decline in recaptures per day was 0.681 for males and 0.667 for females. Gonotrophic cycle duration was estimated to be 4.0 d. Absolute population size was calculated using the Lincoln Index and ranged from 861 to 4,612 males and from 2,187 to 19,739 females. The low proportion of females that reach the age when they are potentially infective suggests that N. neivai has a low biological capacity to serve as a vector and that factors such as high biting rates and opportunistic feeding behavior would be needed to enable Leishmania (Viannia) braziliensis Vianna transmission. This agreed with the epidemiological pattern of ACL in southeastern Brazil that is characterized by low incidence, with isolated cases acquired principally within domiciliary habitats

Pathogenicity and reproductive fitness of Pratylenchus brachyurus on cowpea

The lesion nematode Pratylenchus brachyurus is widespread in cowpea plantations throughout the tropics and sub-tropics. However, the pathogenicity of P. brachyurus on cowpea has been scarcely studied. In this work, it was demonstrated in two glasshouse experiments that an isolate (Pb-20) of P brachyurus was pathogenic to cowpea cv. IPA-206, adversely affecting the plant growth and pod formation and filling. Initial population levels of 5000 and 15 000 nematodes per plant caused reduction of root growth and typical decay of root tissue. The third experiment demonstrated that all six cowpea cultivars selected for evaluation supported reproduction of three isolates of P. brachyurus (Pb-20, Pb-21 and Pb-23) in their roots, although the reproduction factor values obtained indicated that they were dissimilar in their reproductive fitness. Low resistance to R brachyurus was reported for at least one tested cultivar, but apparently of an insufficient degree to be effective for field management of the nematode.

DESCRIPTION OF THE MALE REPRODUCTIVE SYSTEM OF THE HERMIT CRAB CALCINUS TIBICEN (DECAPODA: ANOMURA: DIOGENIDAE)

We describe the male reproductive system of the intertidal hermit crab Calcinus tihicen, with emphasis on the sexual apparatus, spermatophore, and spermatozoa. The crabs were collected on the rocky shore of Praia Grande Beach, Ubatuba, southeastern Brazil. The morphological analysis, based on 30 specimens, was made with the use of a stereomicroscope, an optical microscope, and scanning and transmission electron microscopes. The male reproductive system is composed of a pair of juxtaposed testes, located dorsally in the pleon. From each testis emerges a vas deferens that links it to the exterior by the gonopores. located on the base of the fifth pair of pereiopods. The vas deferens has three macroscopically distinct regions that contain spermatophores in different stages of maturation. The spermatophore morphology is similar to that of other members of Paguroidea, having a distal, nearly spherical ampulla containing spermatozoa; an approximately cylindrical peduncle and a proximal foot connecting the spermatophores. We describe, for the first time, the variability in the spermatophore morphology and size in the three regions of the vas deferens using the type species of the genus Calcinus. The spermatozoa have three main regions (the acrosomal vesicle, the nucleus, and the cytoplasm). The morphological similarity of the male reproductive system of C. tihicen with previously studied species of Diogenidae is an indicative of complex phylogenetic relationships among the members of the genus.

Reproductive aspects of Meloetyphlus fuscatus a meloid beetle cleptoparasite of the bee Eulaema nigrita (Hymenoptera, Apidae, Euglossini)

This study investigated the reproductive biology of the meloid beetle Meloetyphlus fuscatus Waterhouse, a cleptoparasite of Eulaema nigrita nests. New E. nigrita nests had rates of cell parasitism by meloids ranging from 3.7% to 15.8%, while in re-used nests the rate of cell parasitism ranged from 1.4% to 18.7%. The adult parasites were never observed trying to leave the host nests. Both sexes mated more than once. Females had a high fecundity (more than 8,000 eggs), and in most cases, deposited their eggs into the empty, old cells of the host. The triungulins (the first larval instars) hatched from eggs 18-20 days after oviposition and dispersed from the host nest by attaching themselves to males as they emerged. The triungulins most likely transfer to female bees during mating and are transported to the nests of their hosts. Within an attacked cell, the triungulin consumes the bee egg and completes its development by consuming the larval food stored in the cell.

Endometrial claudin-4 and leukemia inhibitory factor are associated with assisted reproduction outcome

Background: Claudin-4 (CLDN4) is one of several proteins that act as molecular mediators of embryo implantation. Recently, we examined immunolabeling of leukemia inhibitory factor (LIF) in the endometrial tissue of 52 IVF patients, and found that LIF staining intensity was strongly correlated with successful pregnancy initiation. In the same set of patients, we have now examined endometrial CLDN4 expression, to see how expression intensity may vary with LIF. We examined CLDN4 in the luteal phase of the menstrual cycle, immediately preceding IVF treatment. Our aim was to compare expression of LIF and CLDN4 in the luteal phase, and document these patterns as putative biomarkers for pregnancy. Methods: Endometrial tissue was collected from women undergoing IVF. Endometrial biopsies were obtained during the luteal phase preceding IVF, and were then used for tissue microarray (TMA) immunolabeling of CLDN4. Previously published LIF expression data were then combined with CLDN4 expression data, to determine CLDN4/LIF expression patterns. Associations between successful pregnancy after IVF and combined CLDN4/LIF expression patterns were evaluated. Results: Four patterns of immunolabeling were observed in the endometrial samples: 16% showed weak CLDN4 and strong LIF (CLDN4(-)/LIF(+)); 20% showed strong CLDN4 and strong LIF (LIF(+)/CLDN4(+)); 28% showed strong CLDN4 and weak LIF (CLDN4(+)/LIF(-)); and 36% showed weak CLDN4 and weak LIF (CLDN4(-)/LIF(-)). Successful implantation after IVF was associated with CLDN4(-)/LIF(+)(p = 0.003). Patients showing this endometrial CLDN4(-)/LIF(+) immunolabeling were also 6 times more likely to achieve pregnancy than patients with endometrial CLDN4(+)/LIF(-) immunolabeling (p = 0.007). Conclusion: The combined immunolabeling expression of CLDN4(-)/LIF(+) in endometrial tissue is a potential biomarker for predicting successful pregnancy in IVF candidates.

Suppression subtractive hybridization analysis reveals expression of conserved and novel genes in male accessory glands of the ant Leptothorax gredleri

Background: During mating, insect males eject accessory gland proteins (Acps) into the female genital tract. These substances are known to affect female post-mating behavior and physiology. In addition, they may harm the female, e. g., in reducing its lifespan. This is interpreted as a consequence of sexual antagonistic co-evolution. Whereas sexual conflict abounds in non-social species, the peculiar life history of social insects (ants, bees, wasps) with lifelong pair-bonding and no re-mating aligns the reproductive interests of the sexes. Harming the female during mating would negatively affect male fitness and sexual antagonism is therefore not expected. Indeed, mating appears to increase female longevity in at least one ant species. Acps are presumed to play a role in this phenomenon, but the underlying mechanisms are unknown. In this study, we investigated genes, which are preferentially expressed in male accessory glands of the ant Leptothorax gredleri, to determine which proteins might be transferred in the seminal fluid. Results: By a suppression subtractive hybridization protocol we obtained 20 unique sequences (USs). Twelve had mutual best matches with genes predicted for Apis mellifera and Nasonia vitripennis. Functional information (Gene Ontology) was available only for seven of these, including intracellular signaling, energy-dependent transport and metabolic enzyme activities. The remaining eight USs did not match sequences from other species. Six genes were further analyzed by quantitative RT-PCR in three life cycle stages of male ants. A gene with carboxy-lyase activity and one of unpredicted function were significantly overexpressed in accessory glands of sexually mature males. Conclusions: Our study is the first one to investigate differential gene expression in ants in a context related to mating. Our findings indicate that male accessory glands of L. gredleri express a series of genes that are unique to this species, possibly representing novel genes, in addition to conserved ones for which functions can be predicted. Identifying differentially expressed genes might help to better understand molecular mechanisms involved in reproductive processes in eusocial Hymenoptera. While the novel genes could account for rapidly evolving ones driven by intra-sexual conflict between males, conserved genes imply that rather beneficial traits might get fixed by a process described as inter-sexual cooperation between males and females.

Allopatric speciation in ticks: genetic and reproductive divergence between geographic strains of Rhipicephalus (Boophilus) microplus

Background: The cattle tick, Rhipicephalus (Boophilus) microplus, economically impact cattle industry in tropical and subtropical regions of the world. The morphological and genetic differences among R. microplus strains have been documented in the literature, suggesting that biogeographical and ecological separation may have resulted in boophilid ticks from America/Africa and those from Australia being different species. To test the hypothesis of the presence of different boophilid species, herein we performed a series of experiments to characterize the reproductive performance of crosses between R. microplus from Australia, Africa and America and the genetic diversity of strains from Australia, Asia, Africa and America. Results: The results showed that the crosses between Australian and Argentinean or Mozambican strains of boophilid ticks are infertile while crosses between Argentinean and Mozambican strains are fertile. These results showed that tick strains from Africa (Mozambique) and America (Argentina) are the same species, while ticks from Australia may actually represent a separate species. The genetic analysis of mitochondrial 12S and 16S rDNA and microsatellite loci were not conclusive when taken separately, but provided evidence that Australian tick strains were genetically different from Asian, African and American strains. Conclusion: The results reported herein support the hypothesis that at least two different species share the name R. microplus. These species could be redefined as R. microplus (Canestrini, 1887) (for American and African strains) and probably the old R. australis Fuller, 1899 (for Australian strains), which needs to be redescribed. However, experiments with a larger number of tick strains from different geographic locations are needed to corroborate these results.

Experimental Infection of Amblyomma aureolatum Ticks with Rickettsia rickettsii

We experimentally infected Amblyomma aureolatum ticks with the bacterium Rickettsia rickettsii, the etiologic agent of Rocky Mountain spotted fever (RMSF). These ticks are a vector for RMSF in Brazil. R. rickettsii was efficiently conserved by both transstadial maintenance and vertical (transovarial) transmission to 100% of the ticks through 4 laboratory generations. However, lower reproductive performance and survival of infected females was attributed to R. rickettsii infection. Therefore, because of the high susceptibility of A. aureola turn ticks to R. rickettsii infection, the deleterious effect that the bacterium causes in these ticks may contribute to the low infection rates (< 1%) usually reported among field populations of A. aureolatum ticks in RMSF-endemic areas of Brazil. Because the number of infected ticks would gradually decrease after each generation, it seems unlikely that A. aureolatum ticks could sustain R. rickettsii infection over multiple successive generations solely by vertical transmission.

Sequential analysis of global gene expression profiles in immature and in vitro matured bovine oocytes: potential molecular markers of oocyte maturation

Background: Without intensive selection, the majority of bovine oocytes submitted to in vitro embryo production (IVP) fail to develop to the blastocyst stage. This is attributed partly to their maturation status and competences. Using the Affymetrix GeneChip Bovine Genome Array, global mRNA expression analysis of immature (GV) and in vitro matured (IVM) bovine oocytes was carried out to characterize the transcriptome of bovine oocytes and then use a variety of approaches to determine whether the observed transcriptional changes during IVM was real or an artifact of the techniques used during analysis. Results: 8489 transcripts were detected across the two oocyte groups, of which similar to 25.0% (2117 transcripts) were differentially expressed (p < 0.001); corresponding to 589 over-expressed and 1528 under-expressed transcripts in the IVM oocytes compared to their immature counterparts. Over expression of transcripts by IVM oocytes is particularly interesting, therefore, a variety of approaches were employed to determine whether the observed transcriptional changes during IVM were real or an artifact of the techniques used during analysis, including the analysis of transcript abundance in oocytes in vitro matured in the presence of a-amanitin. Subsets of the differentially expressed genes were also validated by quantitative real-time PCR (qPCR) and the gene expression data was classified according to gene ontology and pathway enrichment. Numerous cell cycle linked (CDC2, CDK5, CDK8, HSPA2, MAPK14, TXNL4B), molecular transport (STX5, STX17, SEC22A, SEC22B), and differentiation (NACA) related genes were found to be among the several over-expressed transcripts in GV oocytes compared to the matured counterparts, while ANXA1, PLAU, STC1and LUM were among the over-expressed genes after oocyte maturation. Conclusion: Using sequential experiments, we have shown and confirmed transcriptional changes during oocyte maturation. This dataset provides a unique reference resource for studies concerned with the molecular mechanisms controlling oocyte meiotic maturation in cattle, addresses the existing conflicting issue of transcription during meiotic maturation and contributes to the global goal of improving assisted reproductive technology.

Genetic parameters for productive life traits and reproductive efficiency traits at 6 years in Nellore cattle

The objective of the present study was to estimate (co)variance components for length of productive life (LPL) and some alternative reproductive traits of 6-year-old Nellore cattle. The data set contained 57,410 records for age at first calving from Nellore females and was edited to remove animal records with uncertain paternity and cows with just one piece of calving information. Only animals with age at first calving ranging from 23 to 48 months and calving intervals between 11 and 24 months were kept for analysis. LPL and life production ( LP) were used to describe productive life. LPL was defined as the number of months a cow was kept in the herd until she was 6 years old, given that she was alive at first calving and LP was defined as total number of calves in that time. Four traits were used to describe reproductive traits: two breeding efficiencies on original scale were estimated using Wilcox and Tomar functions (BEW and BET, respectively), and two breeding efficiencies transformed (ASBEW and ASBET, respectively), using the function [arcsine (square root (BEi/100))]. Estimates of heritability for measures of LPL and LP were low and ranged from 0.04 to 0.05. Estimates of heritability for breeding efficiencies on original and transformed scales oscillated from 0.18 to 0.32. Estimates of genetic correlations ranged from -0.57 to 0.79 for LPL and other traits and from 0.28 to 0.63 for LP and other traits.

REPRODUCTIVE BIOLOGY IN FEMALES OF AEGLA STRINATII (DECAPODA: ANOMURA: AEGLIDAE)

Females of Aegla strinatii (n = 466) were sampled monthly (September 2003 to September 2005) by means of sieves and traps from Rio das Ostras (24 degrees 38'16.2 '' S; 48 degrees 24'05.2 '' W), at Jacupiranga State Park, South of Sao Paulo State, Brazil. The reproductive period was markedly seasonal (from May to September) encompassing the Austral late autumn through late winter. This is in accordance to the pattern of reproductive period variations in relation to the latitudinal climate variability verified in species of Aegla. The proportion of adult females exhibiting the ovigerous condition was higher in young/small specimens as compared to old/large ones, and suggests the occurrence of senescence in the latter group. Average size at the onset of functional maturity in females was estimated as 16.66 mm of carapace length (rostrum excluded). The number of eggs per ovigerous females ranged from 1 to 325. Eggs are slightly elliptical and average size varied according to embryonic stage. Mean (+/- standard deviation) carapace length of juveniles (n = 118) was 1.50 +/- 0.05 mm (range: 1.40-1.65mm).

The number of reproductive workers in highly eusocial Hymenoptera: monogyny and monandry

Haplodiploidy results in relatedness asymmetries between colony members of highly eusocial Hymenoptera. As a consequence, queen and reproductive workers are more related to their own sons than to each other's male offspring. Kin selection theory predicts multiple optima in male parentage: either the queen or the workers should produce all the males. Nevertheless, shared male parentage is common in highly eusocial hymenopterans. An inclusive fitness model was used to analyze the effect of the number of reproductive workers on male parentage shared by the queen and laying workers by isolating the male component from an inclusive fitness equation using the equal fitness through male condition for each pairwise combination of the three female classes comprised of the queen, laying workers and non-laying workers. The main result of the theoretical analyses showed that the fraction of males produced by workers increases asymptotically with the number of laying workers at an increasingly diminishing rate, tending to an asymptotic value of 0.67. In addition, as the number of laying workers increases, the share of male parentage converges to that of non-laying workers. The diminishing return effect on male parentage share depending on the number of reproductive workers leads us to expect the number of reproductive workers to be relatively small in a stingless bee colony, even in the absence of productivity costs. The available data confirms this hypothesis, as there is an unusually small number of reproductive workers in stingless bee colonies.

REPRODUCTIVE BIOLOGY AND FUNCTIONAL MATURITY IN FEMALES OF AEGLA FRANCA (DECAPODA: ANOMURA: AEGLIDAE)

We describe the reproductive period. fecundity, and average size at the onset of functional maturity of female Aegla franca, the northernmost distributed aeglid species. The reproductive period is markedly seasonal and takes place front May (austral mid-autumn) to August (late winter). Ovigerous females appear quite abruptly in the population by May, and this condition is observed in all adult females sampled regardless of their size. The average size at the onset of functional maturity in females, at which 50% of the females sampled during the reproductive period were considered adults, was 12.75 mm CL. The smallest post-ovigerous female measured 12.06 mm carapace length (CL). Mean fecundity (+/- S.D.) from 41 females bearing early and intermediate eggs was 129.1 +/- 32.2 and corresponded to a mean female CL of 14.11 mm. The elliptical-shaped eggs exhibited significant increase in size along the development stages. The third pair of pleopods bore higher number of eggs than the others. Compiled information regarding the reproductive period reported for aeglids revealed all increase in the breeding period length with latitude. The reproductive period tends to be shorter in localities under larger rainfall variation and smaller temperature variability than in sites with opposite climate conditions. Eggs tend to be fewer in number and larger in size towards lower latitudes. We present an hypothesis that stream water velocity might act as a major selective pressure during the early life history of fluvial aeglids with direct effect on the reproductive pattern.

Molecules Clarify a Cnidarian Life Cycle - The ""Hydrozoan'' Microhydrula limopsicola Is an Early Life Stage of the Staurozoan Haliclystus antarcticus

Background: Life cycles of medusozoan cnidarians vary widely, and have been difficult to document, especially in the most recently proposed class Staurozoa. However, molecular data can be a useful tool to elucidate medusozoan life cycles by tying together different life history stages. Methodology/Principal Findings: Genetic data from fast-evolving molecular markers (mitochondrial 16S, nuclear ITS1, and nuclear ITS2) show that animals that were presumed to be a hydrozoan, Microhydrula limopsicola (Limnomedusae, Microhydrulidae), are actually an early stage of the life cycle of the staurozoan Haliclystus antarcticus (Stauromedusae, Lucernariidae). Conclusions/Significance: Similarity between the haplotypes of three markers of Microhydrula limopsicola and Haliclystus antarcticus settles the identity of these taxa, expanding our understanding of the staurozoan life cycle, which was thought to be more straightforward and simple. A synthetic discussion of prior observations makes sense of the morphological, histological and behavioral similarities/congruence between Microhydrula and Haliclystus. The consequences are likely to be replicated in other medusozoan groups. For instance we hypothesize that other species of Microhydrulidae are likely to represent life stages of other species of Staurozoa.45

Hormone-regulated expression and distribution of versican in mouse uterine tissues

Background: Remodeling of the extracellular matrix is one of the most striking features observed in the uterus during the estrous cycle and after hormone replacement. Versican (VER) is a hyaluronan-binding proteoglycan that undergoes RNA alternative splicing, generating four distinct isoforms. This study analyzed the synthesis and distribution of VER in mouse uterine tissues during the estrous cycle, in ovariectomized (OVX) animals and after 17beta-estradiol (E2) and medroxyprogesterone (MPA) treatments, either alone or in combination. Methods: Uteri from mice in all phases of the estrous cycle, and animals subjected to ovariectomy and hormone replacement were collected for immunoperoxidase staining for versican, as well as PCR and quantitative Real Time PCR. Results: In diestrus and proestrus, VER was exclusively expressed in the endometrial stroma. In estrus and metaestrus, VER was present in both endometrial stroma and myometrium. In OVX mice, VER immunoreaction was abolished in all uterine tissues. VER expression was restored by E2, MPA and E2+MPA treatments. Real Time PCR analysis showed that VER expression increases considerably in the MPA-treated group. Analysis of mRNA identified isoforms V0, V1 and V3 in the mouse uterus. Conclusion: These results show that the expression of versican in uterine tissues is modulated by ovarian steroid hormones, in a tissue-specific manner. VER is induced in the myometrium exclusively by E2, whereas MPA induces VER deposition only in the endometrial stroma.