Molecular Epidemiology of Cryptosporidiosis in Children in Malawi

ABSTRACT: Few studies have examined the molecular epidemiology of cryptosporidiosis in developing countries. In this study, DNA of 69 microscopy-positive human fecal samples collected from Malawi were examined by multilocus genetic analyses. From 43, 27 and 28 of the samples, the SSU rRNA, 70 kDa heat shock protein (HSP70) and 60 kDa glycoprotein (GP60) genes, respectively, were successfully PCR-amplified. Restriction analysis of the SSU PCR products showed that 41 of the 43 PCR-positive samples had C. hominis and 2 had C. parvum. Sequence analysis of the HSP70 and GP60 gene contirmed the species identification by SSU rRNA PCR-RFLP analysis, but also revealed high intraspecific variations. Altogether, six HSP70 subtypes and six GP60 subtypes (belonging to lour subtype alleles) of C. hominis were found. Linkage diseyuilibrum analysis of the two genetic loci showed possible intraspecitic recombination. Thus, cryptosporidiosis in the study area was largely caused by anthroponotic transmission. The high intraspecitic variation and existence of genetic recombination were probably results of high transmission of cryptosporidiosis in this area.     

Fine-Scale Analysis of Mount Graham Red Squirrel Habitat Following Disturbance

ABSTRACT Habitat destruction and degradation are major factors in reducing abundance, placing populations and species in jeopardy. Monitoring changes to habitat and identifying locations of habitat for a species, after disturbance, can assist mitigation of the effects of human-caused or -amplified habitat disturbance. Like many areas in the western United States, the Pinaleño Mountains of southeastern Arizona, USA, have suffered catastrophic fire and large-scale insect outbreaks in the last decade. The federally endangered Mt. Graham red squirrel (Tamiasciurus hudsonicus grahamensis) is only found in the Pinaleño Mountains, and to assess effects of forest disturbance on habitat we modeled their potential habitat by identifying characteristics of cover surrounding their centrally defended middens. We classified high-spatial resolution satellite imagery into ground cover classes, and we used logistic regression to determine areas used by squirrels. We also used known midden locations in conjunction with slope, elevation, and aspect to create a predictive habitat map. Squirrels selected areas of denser forest with higher seedfall for midden sites. Among active middens, those in the densest and least damaged forests were occupied in more seasons than those in more fragmented and damaged areas. The future conservation of red squirrels and the return of healthy mature forests to the Pinaleño Mountains will rely on preservation of mixed conifer zones of the mountain and active restoration of spruce-fir forests to return them to squirrel habitat. Our ability to evaluate the spectrum of fine- to coarse-scale disturbance effects (individual tree mortality to area wide boundaries of a disturbance) with high-resolution satellite imagery shows the utility of this technique for monitoring future disturbances to habitat of imperiled species.     

Net changes in regional woody vegetation cover and carbon storage in Texas Drylands, 1937–1999

Although local increases in woody plant cover have been documented in arid and semiarid ecosystems worldwide, there have been few long‐term, large‐scale analyses of changes in woody plant cover and aboveground carbon (C) stocks. We used historical aerial photography, contemporary Landsat satellite data, field observations, and image analysis techniques to assess spatially specific changes in woody vegetation cover and aboveground C stocks between 1937 and 1999 in a 400‐km² region of northern Texas, USA. Changes in land cover were then related to topo‐edaphic setting and historical land‐use practices. Mechanical or chemical brush management occurred over much of the region in the 1940–1950s. Rangelands not targeted for brush management experienced woody cover increases of up to 500% in 63 years. Areas managed with herbicides, mechanical treatments or fire exhibited a wide range of woody cover changes relative to 1937 (?75% to + 280%), depending on soil type and time since last management action. At the integrated regional scale, there was a net 30% increase in woody plant cover over the 63‐year period. Regional increases were greatest in riparian corridors (33%) and shallow clay uplands (26%) and least on upland clay loams (15%). Allometric relationships between canopy cover and aboveground biomass were used to estimate net aboveground C storage changes in upland (nonriparian) portions of regional landscapes. Carbon stocks increased from 380 g C m^?2 in 1937 to 500 g C m^?2 in 1999, a 32% net increase across the 400 km² region over the 63‐year period. These plant C storage change estimates are highly conservative in that they did not include the substantial increases in woody plant cover observed within riparian landscape elements. Results are discussed in terms of implications for ‘carbon accounting’ and the global C cycle.     

The Life Cycle of Cryptosporidium baileyi n. sp. (Apicomplexa, Cryptosporidiidae) Infecting Chickens

ABSTRACT. The life cycle and morphology of a previously undescribed species of Cryptosporidium isolated from commercial broiler chickens is described. The prepatent period for Cryptosporidium baileyi n. sp. was three days post oral inoculation (PI) of oocysts, and the patent period was days 4–24 PI for chickens inoculated at two days of age and days 4–14 for chickens inoculated at one and six months of age. During the first three days PI, most developmental stages of C. baileyi were found in the microvillous region of enterocytes of the ileum and large intestine. By day 4 PI, most parasites occurred in enterocytes of the cloaca and bursa of Fabricius (BF). Mature Type I meronts with eight merozoites first appeared 12 h PI and measured 5.0 × 4.9 μm. Mature Type II meronts with four merozoites and a large granular residuum first appeared 48 h PI and measured 5.1 × 5.1 μm. Type I meronts with eight short merozoites and a large homogeneous residuum first appeared 72 h PI and measured 5.2 × 5.1 μm. Microgamonts (4.0 × 4.0 μm) produced 16 micro-gametes that penetrated into macrogametes (4.7 × 4.7 μm). Macrogametes gave rise to two types of oocysts that sporulated within the host cells. Most were thick-walled oocysts (6.3 × 5.2 μm), the resistant forms that passed unaltered in the feces. Some were thin-walled oocysts whose wall (membrane) readily ruptured upon release from the host cell. Sporozoites from thin-walled oocysts were observed penetrating enterocytes in mucosal smears. The presence of thin-walled, autoinfective oocysts and the recycling of Type I meronts may explain why chickens develop heavy intestinal infections lasting up to 21 days. Oocysts of C. baileyi were inoculated orally into several animals to determine its host specificity. Cryptosporidium baileyi did not produce infections in suckling mice and goats or in two-dayold or two-week-old quail. One of six 10-day-old turkeys had small numbers of asexual stages only in the BF. Four of six one-day-old turkeys developed mild infections only in the BF, and sexual stages of the parasite were observed in only one of the four. All seven one-day-old ducks and seven two-day-old geese developed heavy infections only in the BF with all known developmental stages present.     

The role of leaf wounding and an epigeal predator on caterpillar damage to tomato plants

The effects of leaf wounding and the presence of a New Zealand carabid predator, Megadromus antarcticus, on the mortality of, and leaf damage caused by, Spodoptera litura on tomatoes were examined in glass houses. The presence of the non-climbing carabid increased S. litura mortality with a subsequent decrease in leaf damage. Leaf wounding produced a possible decrease in herbivory but did not affect S. litura mortality. Neither leaf wounding or the presence of carabids affected the vertical distribution of leaf damage on the tomato plants. The role of M. antarcticus as a biocontrol agent for tomato pests in New Zealand is briefly discussed.     

In Vitro Excystation of Caryospora simplex (Apicomplexa: Eimeriorina)1

In vitro excystation of sporozoites of the heteroxenous coccidian Caryospora simplex Léger, 1904 (Apicomplexa: Eimeriorina) is described. Sporocysts freed mechanically from oocysts released a maximum of 51% of their sporozoites within 45 min at 25°C and a maximum of 74% within 20 min at 37°C when incubated in a 0.25% (w/v) trypsin–0.75% (w/v) sodium taurocholate (bile salt) excystation solution. At emergence from sporocysts, sporozoites were weakly motile then became highly active after about 2 min in excystation solution. Sporozoites within sporocysts exposed to bile salt only became highly motile within 25 min at 25°C and within 15 min at 37°C but did not excyst. When exposed only to trypsin at the above temperatures, the Stieda body dissolved; the substieda body remained intact, and the sporozoites exhibited only limited motility within sporocysts; only a few excysted. Intact, sporulated oocysts incubated at 25° or 37°C in 0.02 M cysteine-HC1 and a 50% CO₂ atmosphere for 18 h had no morphologic changes in the oocyst wall. Further incubation of these intact oocysts in excystation solution for 30 min at 37°C caused neither motility of sporozoites within sporocysts nor excystation. Grinding oocysts for 30 sec in a motor-driven, teflon-coated tissue grinder caused motility of some sporozoites within sporocysts but did not result in excystation.