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.

Determination of the Most Efficient Temperature for Radiofrequency Ablation of Renal Cells: A Prospective Study in Dogs

Background and Purpose: Radiofrequency (RF) ablation of renal tumors is a major technique for tumor cell destruction while preserving healthy renal parenchyma. There is no consensus in the literature regarding the optimal temperature, impedance, and time for RF application for effective cell destruction. This study investigated two variables while keeping time unchanged: Temperature for RF cell destruction and tissue impedance in dog kidneys. Materials and Methods: Sixteen dogs had renal punctures through videolaparoscopy for RF interstitial tissue ablation. A RF generator was applied for 10 minutes to the dog's kidney at different target temperatures: 80 degrees C, 90 degrees C, and 100 degrees C. On postoperative day14, the animals were sacrificed and nephrectomized. All lesions were macroscopically and microscopically examined. The bioelectrical impedance was evaluated at three different temperatures. Results: Renal injuries were wider and deeper at 90 degrees C (P < 0.001), and they were similar at 80 degrees C and 100 degrees C. The bioelectrical impedance was lower at 90 degrees C than at the temperatures of 80 degrees C and 100 degrees C (P < 0.001). Viable cells in the RF ablation tissue area were not found in the microscopic examination. Conclusion: The most effective cell destruction in terms of width and depth was achieved at 90 degrees C, which was also the optimal temperature for tissue impedance. RF ablation of renal cells eliminated all viable cells.

Myeloid differentiation factor 88 is required for resistance to Neospora caninum infection

Neospora caninum is an intracellular parasite that causes major economic impact on cattle raising farms, and infects a wide range of warm-blooded hosts worldwide. Innate immune mechanisms that lead to protection against this parasite are still unknown. In order to investigate whether myeloid differentiation factor 88 (MyD88) is required for resistance against N. caninum, genetically deficient mice (MyD88(-/-)) and wild type littermates were infected with live tachyzoites and the resistance to infection was evaluated. We found that sub-lethal tachyzoite doses induced acute mortality of MyD88(-/-) mice, which succumbed to infection due to uncontrolled parasite replication. Higher parasitism in MyD88(-/-) mice was associated with the lack of IL-12 production by dendritic cells, delayed IFN-gamma responses by NKT, CD4(+) and CD8(+) T lymphocytes, and production of high levels of IL-10. MyD88(-/-) mice replenished with IL-12 and IFN-gamma abolished susceptibility as the animals survived throughout the experimental period. We conclude that protective IFN-gamma-mediated immunity to N. caninum is dependent on initial MyD88 signaling, in a mechanism triggered by production of IL-12 by dendritic cells. Further knowledge on Toll-like receptor recognition of N. caninum antigens is encouraged, since it could generate new prophylactic and therapeutic tools to control parasite burden.

First Report of the Isolation and Molecular Characterization of Rickettsia amblyommii and Rickettsia felis in Central America

During 2010, 15 adult ticks, identified as Amblyomma cajennense, were collected from horses in Cahuita and Turrialba districts, whereas 7 fleas, identified as Ctenocephalides felis, were collected from a dog in San Jose city, Costa Rica. In the laboratory, three A. cajennense specimens, two from Cahuita and one from Turrialba, were individually processed for rickettsial isolation in cell culture, as was a pool of seven fleas. Rickettsiae were successfully isolated and established in Vero cell culture from the three ticks and from a pool of seven fleas in C6/36 cell culture. The three tick isolates were genotypically identified as Rickettsia amblyommii, and the flea isolate was identified as Rickettsia felis through DNA sequencing of portions of the rickettsial genes gltA, ompA, and ompB of each isolate. In addition, other seven ticks were shown to contain rickettsial DNA. Polymerase chain reaction products of at least two of these ticks were sequenced and also showed to correspond to R. amblyommii. Overall, 66.7% (10/15) of the A. cajennense adult ticks were found to be infected with rickettsiae. This is the first report of a successful isolation in cell culture of R. amblyommii and R. felis from Central America.

Rocky Mountain Spotted Fever in Dogs, Brazil

Clinical illness caused by Rickettsia rickettsii in dogs has been reported solely in the United States. We report 2 natural clinical cases of Rocky Mountain spotted fever in dogs in Brazil. Each case was confirmed by seroconversion and molecular analysis and resolved after doxycycline therapy.

Determination and Molecular Analysis of the Complete Genome Sequence of Two Wild-Type Rabies Viruses Isolated from a Haematophagous Bat and a Frugivorous Bat in Brazil

The complete genome sequences of two Brazilian wild-type rabies viruses (RABV), a BR-DR1 isolate from a haematophagous bat (Desmodus rotundus) and a BR-AL1 isolate from a frugivorous bat (Artibeus lituratus), were determined. The genomes of the BR-DR1 and RR-AL1 had 11,923 and 11,922 nt, respectively, and both encoded the five standard genes of rhabdoviruses. The complete nucleotide sequence identity between the BR-DR1 and BR-AL1 isolates was 97%. The BR-DR1 and BR-AL1 isolates had some conserved functional sites revealed by the fixed isolates, whereas both isolates had unique amino acid substitutions in the antigenic region IV of the nucleocapsid gene. Therefore, it is speculated that both isolates were nearly identical in virologic character. According to our phylogenetic analysis based on the complete genomes, both isolates belonged to genotype 1, and to the previously defined ""vampire bat-related RABV lineage"" which consisted of mainly D. rotundus- and A. lituratus- isolates; however, a branch pattern with high bootstrap values suggested that BR-DR1 was more closely related to the 9001FRA isolate, which was collected from a dog bitten by a bat in French Guiana, than to BR-AL1. This result suggests that the vampire bat-related RABV lineage includes Brazilian vampire bat and Brazilian frugivorous bat RABV and is further divided into Brazilian vampire bat and Brazilian frugivorous bat RABV sub-lineages. The phylogenetic analysis based on the complete genomes was valuable in discriminating among very closely related isolates.

Experimental Infection of Rhipicephalus sanguineus Ticks with the Bacterium Rickettsia rickettsii, Using Experimentally Infected Dogs

We evaluated if Rickettsia rickettsii-experimentally infected dogs could serve as amplifier hosts for Rhipicephalus sanguineus ticks. In addition, we checked if Rh. sanguineus ticks that acquired Ri. rickettsii from dogs could transmit the bacterium to susceptible hosts (vector competence), and if these ticks could maintain the bacterium by transstadial and transovarial transmissions. Uninfected larvae, nymphs, and adults of Rh. sanguineus were allowed to feed upon three groups of dogs: groups 1 (G1) and 2 (G2) composed of Ri. rickettsii-infected dogs, infected intraperitoneally and via tick bites, respectively, and group 3 composed of uninfected dogs. After larval and nymphal feeding on rickettsemic dogs, 7.1-15.2% and 35.8-37.9% of the molted nymphs and adults, respectively, were shown by polymerase chain reaction (PCR) to be infected by Ri. rickettsii, confirming that both G1 and G2 dogs were efficient sources of rickettsial infection (amplifier host), resulting in transstadial transmission of the agent. These infected nymphs and adults successfully transmitted Ri. rickettsii to guinea pigs, confirming vector competence after acquisition of the infection from rickettsemic dogs. Transovarial transmission of Ri. rickettsii was observed in engorged females that had been infected as nymphs by feeding on both G1 and G2 dogs, but not in engorged females that acquired the infection during adult feeding on these same dogs. In the first case, filial infection rates were generally <50%. No tick exposed to G3 dogs was infected by rickettsiae in this study. No substantial mortality difference was observed between Ri. rickettsii-infected tick groups (G1 and G2) and uninfected tick group (G3). Our results indicate that dogs can be amplifier hosts of Ri. rickettsii for Rh. sanguineus, although only a minority of immature ticks (<45%) should become infected. It appears that Rh. sanguineus, in the absence of horizontal transmission, would not maintain Ri. rickettsii through successive generations, possibly because of low filial infection rates.

T cells, adhesion molecules and modulation of apoptosis in visceral leishmaniasis glomerulonephritis

Background: Immune complex deposition is the accepted mechanism of pathogenesis of VL glomerulopathy however other immune elements may participate. Further in the present study, no difference was seen between immunoglobulin and C3b deposit intensity in glomeruli between infected and non-infected dogs thus T cells, adhesion molecules and parameters of proliferation and apoptosis were analysed in dogs with naturally acquired VL from an endemic area. The dog is the most important domestic reservoir of the protozoa Leishmania (L.) chagasi that causes visceral leishmaniasis (VL). The similarity of VL manifestation in humans and dogs renders the study of canine VL nephropathy of interest with regard to human pathology. Methods: From 55 dogs with VL and 8 control non-infected dogs from an endemic area, kidney samples were analyzed by immunohistochemistry for immunoglobulin and C3b deposits, staining for CD4+ and CD8+ T cells, ICAM-1, P-selectin and quantified using morphometry. Besides proliferation marker Ki-67, apoptosis markers M30 and TUNEL staining, and related cytokines TNF-alpha, IL-1 alpha were searched and quantified. Results: We observed similar IgG, IgM and IgA and C3b deposit intensity in dogs with VL and non-infected control dogs. However we detected the Leishmania antigen in cells in glomeruli in 54, CD4+ T cells in the glomeruli of 44, and CD8+ T cells in 17 of a total of 55 dogs with VL. Leishmania antigen was absent and T cells were absent/scarse in eight non-infected control dogs. CD 4+ T cells predominate in proliferative patterns of glomerulonephritis, however the presence of CD4+ and CD8+ T cells were not different in intensity in different patterns of glomerulonephritis. The expression of ICAM-1 and P-selectin was significantly greater in the glomeruli of infected dogs than in control dogs. In all patterns of glomerulonephritis the expression of ICAM-1 ranged from minimum to moderately severe and P-selectin from absent to severe. In the control animals the expression of these molecules ranged from absent to medium intensity. It was not observed any correlation between severity of the disease and these markers. There was a correlation between the number of Leishmania antigen positive cells and CD4+ T cells, and between the number of CD4+ T cells and CD8+ T cells. In dogs presenting different histopathological patterns of glomerulonephritis, parameters of proliferation and apoptosis were studied. Ki-67, a proliferative marker, was not detected locally, but fewer apoptotic cells and lower TNF-alpha expression were seen in infected animals than in non-infected controls. Conclusion: Immunopathogenic mechanisms of VL glomerulonephritis are complex and data in the present study suggest no clear participation of immunoglobulin and C3b deposits in these dogs but the possible migration of CD4+ T cells into the glomeruli, participation of adhesion molecules, and diminished apoptosis of cells contributing to determine the proliferative pattern of glomerulonephritis in VL.

SOFT TISSUE INTEGRATION IN THE NECK AREA OF TITANIUM IMPLANTS - AN ANIMAL TRIAL

Dental implant materials are required to enable good apposition of bone and soft tissues. They must show sufficient resistance to chemical, physical and biological stress in the oral cavity to achieve good long-term outcomes. A critical issue is the apposition of the soft tissues, as they have provided a quasi-physiological closure of oral cavity. The present experiment was performed to study the peri-implant tissue response to non-submerged (1-stage) implant installation procedures. Two different implants types (NobelBiocare, NobelReplace (R) Tapered Groovy 4.3 x 10 mm and Replace (R) Select Tapered TiU RP 4.3 x 10 mm) were inserted into the right and left sides of 8 domestic pigs (Sus scrofa domestica) mandibles, between canines and premolars and immediately provided with a ceramic crown. Primary implant stability was determined using ressonance frequency analysis. Soft tissue parameters were assessed: sulcus depth (SDI) and junctional epithelium (JE). Following 70 days of healing, jaw sections were processed for histology and histomorphometric examination. Undecalcified histological sections demonstrated osseointegration with direct bone contact. The soft tissue parameters revealed no significant differences between the two implant types. The peri-implant soft tissues appear to behave similarly in both implant types.

Flow Visualization in Arteriovenous Fistula and Aneurysm Using Computational Fluid Dynamics

The arteriovenous fistula (AVF) is characterized by enhanced blood flow and is the most widely used vascular access for chronic haemodialysis (Sivanesan et al., 1998). A large proportion of the AVF late failures are related to local haemodynamics (Sivanesan et al., 1999a). As in AVF, blood flow dynamics plays an important role in growth, rupture, and surgical treatment of aneurysm. Several techniques have been used to study the flow patterns in simplified models of vascular anastomose and aneurysm. In the present investigation, Computational Fluid Dynamics (CFD) is used to analyze the flow patterns in AVF and aneurysm through the velocity waveform obtained from experimental surgeries in dogs (Galego et al., 2000), as well as intra-operative blood flow recordings of patients with radiocephalic AVF ( Sivanesan et al., 1999b) and physiological pulses (Aires, 1991), respectively. The flow patterns in AVF for dog and patient surgeries data are qualitatively similar. Perturbation, recirculation and separation zones appeared during cardiac cycle, and these were intensified in the diastole phase for the AVF and aneurysm models. The values of wall shear stress presented in this investigation of AVF and aneurysm models oscillated in the range that can both cause damage to endothelial cells and develop atherosclerosis.

Three-Dimensional Stress Distribution in the Human Periodontal Ligament in Masticatory, Parafunctional, and Trauma Loads: Finite Element Analysis

Background: The presence of the periodontal ligament (PDL) makes it possible to absorb and distribute loads produced during masticatory function and other tooth contacts into the alveolar process via the alveolar bone proper. However, several factors affect the integrity of periodontal structures causing the destruction of the connective matrix and cells, the loss of fibrous attachment, and the resorption of alveolar bone. Methods: The purpose of this study was to evaluate the stress distribution by finite element analysis in a PDL in three-dimensional models of the upper central incisor under three different load conditions: 100 N occlusal loading at 45 degrees (model 1: masticatory load); 500 N at the incisal edge at 45 degrees (model 2: parafunctional habit); and 800 N at the buccal surface at 90 degrees (model 3: trauma case). The models were built from computed tomography scans. Results: The stress distribution was quite different among the models. The most significant values (harmful) of tensile and compressive stresses were observed in models 2 and 3, with similarly distinct patterns of stress distributions along the PDL. Tensile stresses were observed along the internal and external aspects of the PDL, mostly at the cervical and middle thirds. Conclusions: The stress generation in these models may affect the integrity of periodontal structures. A better understanding of the biomechanical behavior of the PDL under physiologic and traumatic loading conditions might enhance the understanding of the biologic reaction of the PDL in health and disease. J Periodontol 2009;80:1859-1867.

Inorganic elements in the fat bodies of Diatraea saccharalis (Lepidoptera: Crambidae) larvae parasitized by Cotesia flavipes (Hymenoptera: Braconidae)

Koinobiont parasitoids use several strategies to regulate the host`s physiological processes during parasitism. Although many aspects of host-parasitoid interactions have been explored, studies that attempted to assess the effects of parasitism on the availability of inorganic elements in the host are virtually nonexistent. Therefore, we aimed to evaluate the effects of parasitism on the concentrations of inorganic elements in the fat bodies of larvae of Diatraea saccharalis (Lepidoptera: Crambidae) during the development of the parasitoid Cotesia flavipes (Hymenoptera: Braconidae), by using total reflection X-ray fluorescence (TXRF). TXRF analysis allowed comparisons of the changes in the availability of the elements P. S. K, Ca, Cr, Fe, Ni, Cu, and Zn in the fat body tissues of D. saccharalis larvae parasitized by C. flavipes. Overall, the concentration of inorganic elements was higher early in parasitoid development (1 and 3 days after parasitism) compared to non-parasitized larvae, but much lower towards the end of parasitoid development (7 and 9 days after parasitism). Ca, K, and S were reduced after the fifth day of parasitism, which affected the total abundance of inorganic elements observed in the fat bodies of the parasitized hosts. The regulatory mechanisms or pathological effects related to the observed variation of the host inorganic elements induced by the parasitoid remain unknown, but there might be a strategy to make these elements available to the parasitoid larvae at the end of their development, when higher metabolic activity of the host fat body is required to sustain parasitoid growth. The observed variation of the host`s inorganic elements could also be related to the known effects of parasitism on the host`s immune response. (C) 2010 Elsevier Inc. All rights reserved.

Changes in the hemolymph and fat body metabolites of Diatraea saccharalis (Fabricius) (Lepidoptera : Crambidae) parasitized by Cotesia flavipes (Cameron) (Hymenoptera : Braconidae)

The koinobiont Cotesia flavipes responds to and is influenced by biochemical changes in the host hemolymph composition, Diatraea saccharalis. Changes in the composition of macronutrients may occur due to the hosts own development or by changes induced after parasitization. These changes occur to facilitate parasitoid invasion and to make the host internal environment suitable to parasitoid immature development. Therefore, changes in the availability of stored and circulating nutrients may correlate with the nutritional requirements of specific parasitoid immature stages. In here, we describe changes in the biochemical composition of parasitized and control larvae at different stages of parasitoid development to gain information on C flavipes host regulation and on its quantitative immature nutritional requirements. Total proteins, lipids and carbohydrates free in the hemolymph or stored in host fat bodies, and the SDS-PAGE protein profile of the hemolymph were evaluated in control and parasitized 6th instar during the whole parasitoid development. Changes in the total protein available in the host hemolymph were detected soon after parasitization, but carbohydrate and lipids were observed to differ only towards parasitoid larvae egression. Although C. flavipes affected the availability of all macronutrients observed in the host hemolymph, lipids and proteins stored in the host fat bodies were unaffected. However, carbohydrate concentration at the end of parasitoid larval development was much lower in parasitized than in control larvae at the same stage of development. SDS-PAGE analysis indicated C flavipes up-regulated two host proteins (125 and 48 kDa) and released two parasitism-specific proteins towards the end of parasitoid larval development. We provide a discussion on the role these changes may have on the process of host regulation and their possible requirement to sustain parasitoid development. (C) 2007 Elsevier Inc. All rights reserved.

Resistance of Santa Ines and crossbred ewes to naturally acquired gastrointestinal nematode infections

This trial was carried out in Piracicaba, Sao Paulo State. Brazil. to comparatively evaluate the degree of resistance to naturally acquired gastrointestinal nematode infections in sheep of the following genetic groups purebred Santa Ines (SI), SI crossbred with Dorper (DO x SI), lie de France (IF x SI), Suffolk (SU x SI), and Texel (TE < SI) Fifteen ewes from each group were raised indoors until 12 months of age. At this age, they were moved to pasture that was naturally contaminated by nematode infective larvae and were evaluated from December to May. 2007. Rainfall ranged from 267 mm in January to 37 mm in April Maximum and minimum mean temperatures ranged from 32 5 degrees C to 19 0 degrees C in March and from 25.9 degrees C to 12.8 degrees C in May. There was an increase in the mean number of eggs per gram of feces (EPG) after animals were placed on pasture with significant difference between the SI (80 EPG) and IF x SI (347 EPG) groups in January: and the DO x SI (386 EPG) and TE x SI (258 EPG) groups in May. The highest mean fecal egg count (FEC), 2073 EPG, was recorded for the TE x SI group in February. All groups showed a progressive reduction in body weight throughout the experiment of 12.0% (TE x SI) to 15.9% (SU x SI). In general. the animals with the highest FEC presented the lowest packed cell volumes (PCV): the highest correlation coefficient between FEC x PCV occurred in the SU x SI sheep in January (r = -0.70; P < 0.01). Similarly, there was an inverse relationship between FEC and blood eosinophil Values, with the highest correlation coefficient in the TE x SI sheep in February (r = -0.64; P < 0.05). Immunoglobulin G (IgG) levels against Haemonchus contortus antigens increased in all groups as a result of the exposure to parasites and remained relatively constant until the end of the study, with the exceptions of SU x SI and TE x SI, which showed a rise in IgG levels during the last sampling that coincided with a reduction in mean FEC. In conclusion. crossbreeding Santa Ines sheep with any of the breeds evaluated can result in a production increase and the maintenance of a satisfactory degree of infection resistance, especially against H. contortus and Trichostrongylus colubriformis. the major nematodes detected in this flock. (c) 2009 Elsevier B.V. All rights reserved.

Nitric oxide donor trans-[RuCl([15]aneN(4))NO]2+as a possible therapeutic approach for Chagas` disease

Background and purpose: Benznidazole (Bz) is the therapy currently available for clinical treatment of Chagas` disease. However, many strains of Trypanosoma cruzi parasites are naturally resistant. Nitric oxide (NO) produced by activated macrophages is crucial to the intracellular killing of parasites. Here, we investigate the in vitro and in vivo activities against T. cruzi, of the NO donor, trans-[RuCl([15]aneN(4))NO]2+. Experimental approach: Trans-[RuCl([15]aneN(4))NO]2+ was incubated with a partially drug-resistant T. cruzi Y strain and the anti-proliferative (epimastigote form) and trypanocidal activities (trypomastigote and amastigote) evaluated. Mice were treated during the acute phase of Chagas` disease. The anti-T. cruzi activity was evaluated by parasitaemia, survival rate, cardiac parasitism, myocarditis and the curative rate. Key results: Trans-[RuCl([15]aneN(4))NO]2+ was 10- and 100-fold more active than Bz against amastigotes and trypomastigotes respectively. Further, trans-[RuCl([15]aneN(4))NO]2+ (0.1 mM) induced 100% of trypanocidal activity (trypomastigotes forms) in vitro. Trans-[RuCl([15]aneN(4))NO]2+ induced permanent suppression of parasitaemia and 100% survival in a murine model of acute Chagas` disease. When the drugs were given alone, parasitological cures were confirmed in only 30 and 40% of the animals treated with the NO donor (3.33 mu mol center dot kg-1 center dot day-1) and Bz (385 mu mol center dot kg-1 center dot day-1), respectively, but when given together, 80% of the animals were parasitologically cured. The cured animals showed an absence of myocarditis and a normalisation of cytokine production in the sera. In addition, no in vitro toxicity was observed at the tested doses. Conclusions and implications: These findings indicate that trans-[RuCl([15]aneN(4))NO]2+ is a promising lead compound for the treatment of human Chagas` disease. This article is commented on by Machado et al., pp. 258-259 of this issue. To view this commentary visit http://dx.doi.org/10.1111/j.1476-5381.2010.00662.x and to view a related paper in this issue by Silva et al. visit http://dx.doi.org/10.1111/j.1476-5381.2010.00524.x.