Use of PCR-RFLP to identify Leishmania species in naturally-infected dogs

Tissue imprints on Giemsa stained slides from dogs were used to investigate the presence of Leishmania amastigotes by either optical microscopy (OM) or Polymerase chain reaction (PCR) detection of DNA. Samples from skin, spleen, lymph node, liver and bone marrow from a Leishmaniasis endemic area dogs where Leishmania (Leishmania) chagasi and Leishmania (Viannia) braziliensis are sympatric were studied. Dogs were initially diagnosed by Indirect Immunofluorescence (IIF), as which 39 were IIF positive (≥1:40) and 16 negative. The IIF positive dogs were clinically grouped as symptomatic (n = 15), oligosymptomatic (n = 12) and asymptomatic (n = 12). Although PCR positivity was higher in symptomatic dogs, specially their skin samples, there was no significant difference among clinical groups or organs examined. Ten (62.5%) out of 16 IIF and OM negative animals were positive for PCR in at least one organ. Forty-eight positive PCR amplicons were further submitted to RFLP for Leishmania identification. All dogs were infected with L. (L.) chagasi except one, infected with L. (V.) braziliensis. PCR was more efficient than IIF and OM to diagnose canine visceral Leishmaniasis (CVL), regardless of the organ examined and the clinical form present. The use of PCR together with serology helps determining the extension of sub clinical infection in CVL endemic areas and provides a better estimate of the number of dogs to be targeted for control measures. In conclusion, our data reinforce the need for a specific diagnosis of canine infection in areas where diverse Leishmania species are sympatric and demonstrate that PCR–RFLP can be used to identify Leishmania species in dog tissue imprint stained slides.     

Phytoestrogen treatment induces testis alterations in dogs. Potential use in population control

Dog overpopulation is considered a human health risk; they are the terrestrial vector of rabies and reservoirs for other human diseases. Surgical neutering and intratesticular injections have been used in male dogs. Physiological and morphological alterations in reproductive organs can be induced by phytoestrogens. Our goal was to evaluate the effect of oral coumestrol on dog ejaculates and testis histology. Two groups of 5 healthy adult dogs were used. One coumestrolcontaining biscuit was given once a week for a 4 week period to the experimental group. Ejaculates were obtained and evaluated. After treatment, testis were obtained and processed for histology. Compared to controls, treated dogs have reduced tubules (462 ± 1.4 vs 336 ± 2 μ²), spermatogenic epithelium (49.1 ± 0.01 vs 13.3 ± 0.01 μ²), and lumen opening (891 ± 1.4 vs 530 ± 26.9 μ). Ejaculates from treated animals have increased numbers of abnormal spermatozoa and reduced sperm concentration.     

Protected areas do not fulfil the wintering habitat needs of the trans-Saharan migratory Montagu’s harrier

Populations of migratory birds can be affected by events happening at both breeding and wintering grounds. The Sahel is a vast region holding a large number of wintering trans-Saharan migratory European birds, and current land-use changes there may represent a threat for these species. We used satellite tracking data from the migratory Montagu’s harrier to evaluate habitat use of the species during the wintering season, and whether the current network of protected areas is effective to provide their habitat needs during that season. We also developed an ecological niche model for the species in Western-Central Sahel to check if the most suitable sites are included within current protected areas. Tracked harriers occupied a large region encompassing a total of eight countries. The most preferred habitats during winter were croplands and some natural vegetation habitats, especially grasslands. Protected areas only covered a small proportion of the overall wintering grounds of tracked harriers and the most suitable areas for the species in Western-Central Sahel. Increasing the extent of preferred natural habitats within protected areas should benefit the conservation of this and probably other insect-eating raptors. However, substantial increases in extent and number of protected areas in sub-Saharan Africa are very unlike to occur. Conservation actions in the region should therefore be mostly focused on improving land use planning and management outside protected areas, specially enhancing agricultural practices to make biodiversity conservation compatible with poverty alleviation. These can be chiefly targeted at an area of <20,000 km² of very suitable habitat for these species.     

Colombia and Venezuela 1992 wild potato (Solanum sect. Petota) germplasm collecting expedition: taxonomy and new germplasm resources

Summary We conducted a joint Colombia/United States/Venezuela wild potato (Solanum sect. Petota Dumort.) germplasm collecting expedition in Colombia from June 27–August 24, and in Venezuela from August 17–September 15, 1992. The goals of the expedition were to collect germplasm and study the species boundaries of all of the 23 Colombian and Venezuelan taxa accepted by current taxonomists. We made 128 collections of 16 of these taxa, 96 as true seed collections. We collected the first available germplasm collections of S. cacetanum, S. cuatrecasasii, S. estradae, S. lobbianum, S. orocense, S. paramoense, and S. sucubunense, and obtained germplasm collections of S. neovalenzuelae and S. pamplonense as a germplasm exchange from the Colombian national germplasm collection. We had problems identifying some of our collections, and currently are investigating them for species status and interrelationships. We summarize the state of germplasm collections for Colombia and Venezuela, provide our field data regarding the taxonomy of Colombian and Venezuelan wild potatoes, and provide recommendations for future collecting.     

Regional habitat conservation priorities for the American crocodile

The American crocodile is widely distributed in coastal and lowland wetlands in the northern Neotropics. As a result of commercial skin hunting in the 20th century, populations were greatly diminished, but in many areas have initiated a period of recovery since hunting and trade controls were enacted in the 1980s and 1990s. While a great deal of attention has been devoted to regulated commercial use as a management strategy for recovering crocodilian populations, these approaches are limited in their efficacy to deal with issues of habitat loss and fragmentation. Because habitat limitations are expected to be the most critical issue for crocodile conservation in the 21st century, there is an unfulfilled need for alternative strategies that prioritize habitat conservation. Here, we present results of an international effort to identify and prioritize the most critical habitats for this wide ranging species. We quantified information of a group of American crocodile experts and classified 69 areas in eight distinct crocodile bioregions as Crocodile Conservation Units (CCU), the most important areas for the conservation of this species. The relative importance of the CCUs in each bioregion was quantified using an algorithm that weighted factors that the experts considered to be most important for the long term conservation of viable populations of crocodiles. This effort is the initial step in the development of a regional conservation plan for the American crocodile. We identified two bioregions in particular where the creation of protected areas should be given a high priority, the Dry Pacific South America (northern Peru and southern Ecuador) and the Northwest and Central Pacific Mexico.     

Quambalaria eucalypti found on Eucalyptus in Indonesia

The Eucalyptus plantation industry in Indonesia has expanded rapidly during the last few decades. During routine nursery disease surveys, symptoms of a leaf and shoot blight disease were detected on Eucalyptus mother plants. Isolates were obtained from symptomatic tissues and identified using DNA sequence analyses. Phylogenetic analyses showed that the isolates were those of Quambalaria eucalypti. Pathogenicity tests were conducted with isolates of Q. eucalypti on clones of E. pellita and E. grandis × E. pellita hybrids. These resulted in symptoms similar to those observed on naturally infected plants. Eucalyptus genotypes tested showed variation in their susceptibility, highlighting the potential to select and breed for resistance and thus to manage future outbreaks of the disease. This is the first report of the pathogen in Indonesia as well as in Southeast Asia.     

Identification,pathogenicity and control of powdery mildew caused by Erysiphe elevata on Eucalyptus pellita in Sumatra,Indonesia

Powdery mildews are biotrophic ascomycetes that do not typically kill their hosts, but instead interfere with normal plant growth. In Indonesia, white powdery spots were observed forming on the upper part of fully developed leaves of Eucalyptus pellita planted in sand beds of a clonal nursery. Thus, this study aimed to identify the causal agent of eucalypt powdery mildew in Indonesia as well as to develop control strategies to better manage the disease. Pathogen identification was based on morphology, molecular characterization of DNA sequencing the ITS region and pathogenicity test. In addition, considering that some powdery mildews are inhibited by free moisture on leaves, efficacy of foliar watering was tested for disease control. Phylogenetic analyses showed that isolates collected from infected eucalypt plants grouped together with other sequences of the Erysiphe elevata clade with bootstrap support of 99%; a similar result was also confirmed by morphological observations. Pathogenicity of powdery mildew on eucalypt plants was confirmed through artificial inoculation, thus fulfilling the Koch's postulates. For disease control, foliar watering reduced the incidence of powdery mildew on mother plants. The reduction of the disease incidence began to be more evident at 2 weeks after trial establishment, and in the fourth week, the incidence was below 10% in all three treatments, while the control was above 20%. Therefore, we concluded that E. elevata is the causal agent of powdery mildew in Indonesia and that one application of foliar watering per week was enough to minimize the disease impact in the nursery.     

Aversive effect of tannic acid on drinking behavior in mice of an inbred strain: potential animal model for assessing astringency

Astringency, an orosensory sensation associated with dietary tannins, contributes to food appetitiveness/aversiveness. However, astringency perception varies greatly among individuals. This study examined whether genetically homogeneous na?ve mice display appetitiveness/aversiveness when provided with tannin-containing drink solutions. Ingestion of serial dilutions of tannic acid (TA) by inbred mice (A/Snell) was assessed by a one-bottle preference test. Drink intake was far predominant at night (circadian rhythm). TA concentration-dependently inhibited daily drink consumption. Overnight consumption of TA solutions (range = 0.5-8 g/L) decreased linearly to zero during the first night and was recovered significantly during subsequent nights. TA also inhibited drink consumption in another two inbred mouse strains. The protein fraction of saliva collected from naive mice was markedly reactive with TA at the concentrations shown to affect drink consumption. Thus, testing for drink ingestion in inbred mice during short-term (overnight) exposure to tannin-containing liquid foods represents an advantageous animal model for assessing astringency.