Geographical distribution of Amblyomma cajennense (sensu lato) ticks (Parasitiformes: Ixodidae) in Brazil, with description of the nymph of A-cajennense (sensu stricto)

Background: Until recently, Amblyomma cajennense (Fabricius, 1787) was considered to represent a single tick species in the New World. Recent studies have split this taxon into six species. While the A. cajennense species complex or A. cajennense (sensu lato) (s.l.) is currently represented by two species in Brazil, A. cajennense (sensu stricto) (s.s.) and Amblyomma sculptum Berlese, 1888, their geographical distribution is poorly known. Methods: The distribution of the A. cajennense (s.l.) in Brazil was determined by morphological examination of all lots of A. cajennense (s.l.) in two large tick collections of Brazil, and by collecting new material during three field expeditions in the possible transition areas between the distribution ranges of A. cajennense (s.s.) and A. sculptum. Phylogenetic analysis inferred from the ITS2 rRNA gene was used to validate morphological results. Morphological description of the nymphal stage of A. cajennense (s.s.) is provided based on laboratory-reared specimens. Results: From the tick collections, a total 12,512 adult ticks were examined and identified as 312 A. cajennense (s.s.), 6,252 A. sculptum and 5,948 A. cajennense (s.l.). A total of 1,746 ticks from 77 localities were collected during field expeditions, and were identified as 249 A. cajennense (s.s.), 443 A. sculptum, and 1,054 A. cajennense (s.l.) [these A. cajennense (s.l.) ticks were considered to be males of either A. cajennense (s.s.) or A. sculptum]. At least 23 localities contained the presence of both A. cajennense (s.s.) and A. sculptum in sympatry. DNA sequences of the ITS2 gene of 50 ticks from 30 localities confirmed the results of the morphological analyses. The nymph of A. cajennense (s.s.) is morphologically very similar to A. sculptum. Conclusion: Our results confirmed that A. cajennense (s.l.) is currently represented in Brazil by only two species, A. cajennense (s.s.) and A. sculptum. While these species have distinct distribution areas in the country, they are found in sympatry in some transition areas. The current distribution of A. cajennense (s.l.) has important implications to public health, since in Brazil A. sculptum is the most important vector of the bacterium Rickettsia rickettsii, the etiological agent of Brazilian spotted fever

Interacting effects of pollination, water and nutrients on fruit tree performance.

Pollination is critical to fruit production, but the interactions of pollination with plant resources on a plant's reproductive and vegetative features are largely overlooked. We examined the influences of pollination, irrigation and fertilisation on the performance of almond, Prunus dulcis, in northern California. We used a full-factorial design to test for the effects of pollination limitation on fruit production and foliage variables of whole trees experiencing four resource treatments: (i) normal water and nutrients, (ii) reduced water, (iii) no nutrients, and (iv) reduced water and no nutrients. In each of these combinations, we applied three pollination treatments: hand-cross pollination, open-pollination and pollinator exclusion. Pollination strongly affected yield even under reduced water and no nutrient applications. Hand-cross pollination resulted in over 50% fruit set with small kernels, while open-pollinated flowers showed over 30% fruit set with moderate-sized kernels. Pollinator-excluded flowers had a maximum fruit set of 5%, with big and heavy kernels. Reduced water interacted with the open- and hand-cross pollination treatments, reducing yield more than in the pollinator exclusion treatment. The number of kernels negatively influenced the number of leaves, and reduced water and no nutrient applications interacted with the pollination treatments. Overall, our results indicate that the influences of pollination on fruit tree yield interact with the plant availability of nutrients and water and that excess pollination can reduce fruit quality and the production of leaves for photosynthesis. Such information is critical to understand how pollination influences fruit tree performance.

Amblyomma dissimile Koch (Acari: Ixodidae) attacking Primolius maracana Vieillot (Psittaciformes: Psittacidae) in the Amazon region, State of Pará, Brazil.

The tick Amblyomma dissimile Koch feeds preferentially on reptiles (Squamata), although amphibians (Anura) also seem to be important hosts. We report an A. dissimile nymph infesting a blue-winged macaw, Primolius maracana, held in captivity in the Mangal das Garças Park, State of Pará, Brazil. Environmental observations suggest that free-living iguanas (Iguana iguana), which used to walk on the bird enclosure in the park, were the source of the A. dissimile tick that infested the blue-winged macaw. We provide the second world record of a bird host for A. dissimile, and the first bird record for this species in South America.

Amblyomma dissimile Koch (Acari: Ixodidae) attacking Primolius maracana Vieillot (Psittaciformes: Psittacidae) in the Amazon region, state of Pará, Brazil

The tick Amblyomma dissimile Koch feeds preferentially on reptiles (Squamata), although amphibians (Anura) also seem to be important hosts. We report an A. dissimile nymph infesting a blue-winged macaw, Primolius maracana, held in captivity in the Mangal das Garças Park, State of Pará, Brazil. Environmental observations suggest that free-living iguanas (Iguana iguana), which used to walk on the bird enclosure in the park, were the source of the A. dissimile tick that infested the blue-winged macaw. We provide the second world record of a bird host for A. dissimile, and the first bird record for this species in South America.

Fatal cytauxzoonosis in captive-reared lions in Brazil.

Fatal cytauxzoonosis is described in a captive reared lioness (Panthera leo) and its 6-month-old cub. Clinical signs in the lioness included loss of weight, depression, anaemia, loss of hair, dark discolored urine, tachypnoea, nystagmus, deaphness and staggering gait. The cub died after a short period of depression. In the lioness, laboratory examination revealed normochromic normocytic anaemia, neutrophilia, lymphopenia, monocytosis, eosinopenia, thrombocytopenia, proteinuria, pyuria, haematuria and increased. At necropsy the lioness showed marked pulmonary edema and slight gelatinous translucent edema in the mediastinum, petechiae and echymosis disseminated in the serosae, and the intestinal content was red and semiliquid. The cub presented hemothorax, endocardial and pulmonary edema, petechiae in the cardiac serosae, hepatic and splenic congestion and segments of the small intestine with blood stained fluid contents and reddish mesenteric lymph nodes. Histopathological examination of liver, spleen, heart, lungs, intestines, pancreas, mesenteric lymph nodes, kidneys, skeletal muscle, brain and skin revealed large number of intravascular macrophages with their cytoplasm filled with various schizogonic stages of a Theileriidae. Electron microscopy confirmed the presence of schizonts in endothelial-associated macrophages. The diagnosis was established by the finding of the pathognomonic schizonts in macrophages within blood vessels in several organs and tissues from both lions. This is the first report of feline cytauxzoonosis in P. leo and of a confirmed infection by Cytauxzoon felis in felidae in South America.

Evaluation of the mussel Perna perna as a biomonitor of polycyclic aromatic hydrocarbon (PAH) exposure and effects.

The mussel Perna perna was used to monitor and assess areas suspected of oil contamination by transplanting animals from unimpacted to impacted sites and vice versa. The deployment lasted for three months during which total PAH were measured in seawater samples collected every 15 days. PAH concentrations were determined in mussel tissue monthly. The lysosomal neutral red assay and the condition index were used as biomarkers of the adverse biological effects. The PAH concentration in mussels at unimpacted sites, prior to deployment was ca. 100 microg kg(-1) but increased to 300 microg kg(-1) after three months at the contaminated site. Mussels transplanted to the uncontaminated site showed a decrease from 380 to 80 microg kg(-1). Significant changes in lysosomal stability occurred within one month of the beginning of the transposition; in contrast condition indexes were not correlated with the observed changes in PAH concentration.

Polycyclic aromatic hydrocarbon in inter-tidal mussel Perna perna: space-time observations, source investigation and genotoxicity.

The investigation aimed primarily at understanding the PAH record in inter-tidal mussel tissues and evaluating the bivalve performance as a bioindicator for oil contamination. The species Perna perna was used as test organism since it is abundantly distributed in coastal areas of the Americas, Africa and elsewhere. The study was carried out in Guanabara Bay and comprised two observation phases: phase one included seasonal sampling in 8 sites, whereas in phase two 4 sites were examined over 4 years. Among the 35 determined PAH (60-90 microg kg(-1) up to 4000-6000 microg kg(-1)) alkylated homologues predominated by more than 80%. The PAH profile in inter-tidal mussels is largely petrogenic with high contribution of dibenzothiophenes (DBT) and phenanthrenes (Ph). The prevailing petrogenic fingerprint, confirmed by diagnostic ratios, is linked to the properties of the inter-tidal habitat, which favors exposure to oil films. C2DBT/C2Ph and C3DBT/C3Ph ratios, however, show a wide range of values uncorrelated to specific oils. Micronucleus frequencies are significantly related to PAH concentrations, especially to those of alkylated homologues. Genotoxic expression appears at concentrations as low as 300 microg kg(-1) summation operator 35 PAH.

Molecular characterization of Hepatozoon sp. from Brazilian dogs and its phylogenetic relationship with other Hepatozoon spp.

To characterize phylogenetically the species which causes canine hepatozoonosis at two rural areas of Rio de Janeiro State, Brazil, we used universal or Hepatozoon spp. primer sets for the 18S SSU rRNA coding region. DNA extracts were obtained from blood samples of thirteen dogs naturally infected, from four experimentally infected, and from five puppies infected by vertical transmission from a dam, that was experimentally infected. DNA of sporozoites of Hepatozoon americanum was used as positive control. The amplification of DNA extracts from blood of dogs infected with sporozoites of Hepatozoon spp. was observed in the presence of primers to 18S SSU rRNA gene of Hepatozoon spp., whereas DNA of H. americanum sporozoites was amplified in the presence of either universal or Hepatozoon spp.-specific primer sets; the amplified products were approximately 600bp in size. Cloned PCR products obtained from DNA extracts of blood from two dogs experimentally infected with Hepatozoon sp. were sequenced. The consensus sequence, derived from six sequence data sets, were blasted against sequences of 18S SSU rRNA of Hepatozoon spp. available at GenBank and aligned to homologous sequences to perform the phylogenetic analysis. This analysis clearly showed that our sequence clustered, independently of H. americanum sequences, within a group comprising other Hepatozoon canis sequences. Our results confirmed the hypothesis that the agent causing hepatozoonosis in the areas studied in Brazil is H. canis, supporting previous reports that were based on morphological and morphometric analyses.

Diagnosis of Hepatozoon spp. in Amblyomma ovale and its experimental transmission in domestic dogs in Brazil.

Transmission of Hepatozoon spp. to dogs was investigated using four species of ixodid ticks: Rhipicephalus sanguineus, Amblyomma aureolatum, Amblyomma ovale and Amblyomma cajennense. We collected completely or partially engorged adult ticks of these species from dogs that were naturally infested and positive for Hepatozoon spp. We selected some of these ixodids and inoculated them orally in four negative dogs. The other ticks were dissected and examined for oocysts. Of all dogs inoculated orally with R. sanguineus, A. aureolatum, A. cajennense and A. ovale, only the animal that received the macerate of A. ovale was positive; evidence (gametocytes in peripheral blood) of infection was found 63 days after inoculation. Among all dissected ticks, we found only two oocysts; these were similar to those of Hepatozoon canis, and both were recovered from a single A. ovale specimen. We inoculated sporozoites recovered from the oocysts intraperitoneally into a Hepatozoon spp. negative dog, and circulating gametocytes were detected 84 days later. Our study demonstrated that A. ovale can be a vector of Hepatozoon spp. in Brazil.

Imposex and surface sediment speciation: a combined approach to evaluate organotin contamination in Guanabara Bay, Rio de Janeiro, Brazil.

An integrated biological-chemical survey of organotin compounds was carried out in Guanabara Bay, the second most important Brazilian harbor complex. The biological survey revealed high levels of imposex in Stramonita haemastoma populations. Inside the bay values of relative penis length index from 42.7 to 107.6 and vas deferens sequence index from III to VI were found, while organisms collected outside the bay had values ranging from <0.1 to 35.2 and from 0 to II. None of the females sampled inside the bay were normal and imposex was found in all stations. Surface sediments in the bay are contaminated by tributyltin (10-522 ng/g d.w.) and triphenyltin (<3.9-39.4 ng/g d.w.) with greater concentrations close to shipyards and marinas. The observed predominance of parent compounds (TBT and TPT) is commonly found when recent inputs occur, but may also indicate slow degradation processes in the anoxic conditions of these sediments. To our knowledge, this is the first report of a combined imposex-sediment approach to evaluate organotin contamination in marine environments of South America, and also the first report of TPT detection in environmental matrices in this region.

Morphological and neurotoxicological findings in tropical freshwater fish (Astyanax sp.) after waterborne and acute exposure to water soluble fraction (WSF) of crude oil.

The water-soluble fraction (WSF) of crude oil is a complex highly volatile and toxic mixture of hydrocarbon chains (polyaromatics, heterocyclics), phenols, and heterocyclic compounds containing nitrogen and sulfur. To evaluate the toxic effects of WSF in tropical freshwater teleosts and to develop methodologies that could investigate the toxic mechanisms of WSF in tropical organisms, an acute toxicity experiment was conducted with Astyanax sp. Three dilutions (15%, 33%, and 50%) of WSF obtained from Campos Bay's crude oil (Brazil) were used to study morphological and biochemical responses of the fish. Prior to exposure, the distribution and rate of volatilization of the WSF into each aquarium for the same exposure period was quantified by spectrofluorimetry. Five individuals of Astyvanax sp. were exposed to duplicate WSF of 0, 15, 33, and 50% for each of 12-, 24-, and 96-h exposures for a total of 120 individuals. Liver and gills were sampled from five fish from each treatment and were analyzed by histology, scanning and transmission electron microscopy. A fragment of muscle was also collected from each fish to measure acetylcholinesterase activity. Water analysis showed that only 4 h after dilution, an important loss of hydrocarbons in 33% and 50% of WSF was observed. In addition, 50% of hydrocarbon mass was lost in all tested dilutions after 24 h with significant difference for the 50% WSF at all measured times, demonstrating the high volatility of WSF in freshwater. Damage in the liver and the gills included the presence of necrosis, loss of hepatocytes limit, inflammation areas, cellular proliferation, aneurysms, and disorganization of the second lamellae. The 33% WSF significantly reduced acetylcholinesterase activity in fish. Our study demonstrated that the WSF of crude oil caused damage in organs and tissues of tropical freshwater Astyanax sp. and provided also the basis for a better understanding of the toxic mechanisms of WSF in freshwater fishes.

Can B cell epitopes of 60 kDa Ro distinguish systemic lupus erythematosus from Sjögren's syndrome?

Antibodies binding components of the Ro/La (or SSA/SSB) ribonucleoprotein particle are found in the sera of patients with systemic lupus erythematosus (SLE) and Sjögren's syndrome (SS) as well as mothers who give birth to babies with neonatal lupus. Anti-La occurs in a subset of sera that contain anti-Ro, and anti-La is found more commonly in sera of patients with SS than in sera from SLE patients. The fine specificity of autoantibodies binding 60 kDa has been studied extensively. Recent data have suggested that there are disease-specific epitopes which identify patients with either SLE or SS. Alternatively, other data suggest that the B cell epitopes of 60kDa Ro vary according to the presence of anti-La. The present study was undertaken to determine whether binding of putative disease-specific 60 kDa Ro epitopes is associated with the diagnosis of SLE vs SS, or instead associated with the presence of anti-La. Anti-60 kDa Ro positive sera from 24 SLE patients and 44 SS patients were studied for antibodies binding two epitopes of 60 kDa Ro. We find the epitope defined by residues 171-190 is associated with anti-60 kDa Ro without anti-La, regardless of diagnosis. Meanwhile, binding of the epitope defined by residues 215-232 is not commonly found in anti-60 kDa Ro sera, especially in those sera with both anti-60 kDa Ro and anti-La. Thus, the fine specificity of antibody binding to 60 kDa Ro varies according to the presence of anti-La, not to the diagnosis of either SLE or SS.

Is spiritual healing a valid and effective therapy?

This paper briefly reviews the evidence supporting the reality of healing as a scientifically-attested phenomenon, the techniques and mechanisms of healing and the potential value of this therapy if it were developed and integrated into the medical services. The need for further extensive research is indicated and strategies for such research are suggested.

Inhibition of mammalian digestive disaccharidases by polyhydroxy alkaloids.

Several polyhydroxy alkaloids, including the eight presently known to occur in plants, have been compared as inhibitors of mouse gut digestive disaccharidases. The indolizidine castanospermine inhibited all activities tested, but others showed a selectivity which could be of value in studies of carbohydrate digestion and errors of metabolism.

Intestinal absorption of glucose in rats after continuous challenge with larvae of Nippostrongylus brasiliensis.

The rate of glucose absorption and metabolism and the associated fluid transfer of the small intestine of rats was measured in vitro using an everted sac technique after daily doses of 0, 5, 25 or 225 infective larvae of Nippostrongylus for periods of one to five weeks. Though there were occasional significant changes in the physiology of the intestine there were no obvious patterns or trends with respect to time at any infection level. This differs from the response after a single infection. The possible role of immune adaptation and reduced immunogenicity of the worms during repeated infections is considered.

Intestinal glucose absorption in rats after secondary infections with Nippostrongylus brasiliensis.

A challenge infection of Nippostrongylus brasiliensis in immune rats resulted in an earlier onset of intestinal glucose malabsorption and increased glucose metabolism compared with rats receiving a primary infection. Intestinal absorption and metabolism recovered to control levels earlier during a secondary infection. The pattern of changes in absorption and metabolism was probably related to host immunological activity.