Sodalis glossinidius and Wolbachia infections in wild population of Glossina morsitans submorsitans caught in the area of Lake Iro in the south of Chad.

Investigations on the bacterial fauna and their association with trypanosome infections in tsetse fly have revealed contrasting results. This study aimed to detect Wolbachia and S. glossinidius in wild populations of G. m. submorsistans and subsequently, understand the influence that these bacteria may have on the vectorial competence of this tsetse species. Tsetse flies were captured in the area of Lake Iro in the south of Chad using biconical traps. After DNA extraction from each tsetse fly, Sodalis glossinidius and Wolbachia were detected using specific primers. Sodalis glossinidius and Wolbachia infection rates were compared and association studies involving trypanosome infections and S. glossinidius or Wolbachia were performed. From 345 G. m. submorsitans analyzed, 9.0% and 14.5% were respectively infected with S. glossinidius and Wolbachia. Only 2.31% of all tsetse flies were co-infected by the 2 bacteria. Of all trypanosome-infected flies, 7.1% and 9.8% harbored, respectively, S. glossinidius and Wolbachia. No association was observed between Wolbachia and trypanosome infections while a significant association (r = 4.992; P = 0.025) was found between S. glossinidius and the presence of trypanosomes. A significant association (r = 3.147; P = 0.043) was also observed between S. glossinidius and T. simiae; and none with T. congolense or T. godfreyi. This study revealed S. glossinidius and Wolbachia in G. m. submorsitans of the area of lake Iro. It showed that co-infections between Wolbachia and S. glossinidius are rare in wild populations of G. m. submorsitans and that the tripartite associations vary according to trypanosome species as well as symbiotic mricroorganisms.

Diversity of trypanosomes in humans and cattle in the HAT foci Mandoul and Maro, Southern Chad-A matter of concern for zoonotic potential?

BACKGROUND: African trypanosomes are parasites mainly transmitted by tsetse flies. They cause trypanosomiasis in humans (HAT) and animals (AAT). In Chad, HAT/AAT are endemic. This study investigates the diversity and distribution of trypanosomes in Mandoul, an isolated area where a tsetse control campaign is ongoing, and Maro, an area bordering the Central African Republic (CAR) where the control had not started. METHODS: 717 human and 540 cattle blood samples were collected, and 177 tsetse flies were caught. Trypanosomal DNA was detected using PCR targeting internal transcribed spacer 1 (ITS1) and glycosomal glyceraldehyde-3 phosphate dehydrogenase (gGAPDH), followed by amplicon sequencing. RESULTS: Trypanosomal DNA was identified in 14 human samples, 227 cattle samples, and in tsetse. Besides T. b. gambiense, T. congolense was detected in human in Maro. In Mandoul, DNA from an unknown Trypanosoma sp.-129-H was detected in a human with a history of a cured HAT infection and persisting symptoms. In cattle and tsetse samples from Maro, T. godfreyi and T. grayi were detected besides the known animal pathogens, in addition to T. theileri (in cattle) and T. simiae (in tsetse). Furthermore, in Maro, evidence for additional unknown trypanosomes was obtained in tsetse. In contrast, in the Mandoul area, only T. theileri, T. simiae, and T. vivax DNA was identified in cattle. Genetic diversity was most prominent in T. vivax and T. theileri. CONCLUSION: Tsetse control activities in Mandoul reduced the tsetse population and thus the pathogenic parasites. Nevertheless, T. theileri, T. vivax, and T. simiae are frequent in cattle suggesting transmission by other insect vectors. In contrast, in Maro, transhumance to/from Central African Republic and no tsetse control may have led to the high diversity and frequency of trypanosomes observed including HAT/AAT pathogenic species. Active HAT infections stress the need to enforce monitoring and control campaigns. Additionally, the diverse trypanosome species in humans and cattle indicate the necessity to investigate the infectivity of the unknown trypanosomes regarding their zoonotic potential. Finally, this study should be widened to other trypanosome hosts to capture the whole diversity of circulating trypanosomes.

Diversity of tsetse flies and trypanosome species circulating in the area of Lake Iro in southeastern Chad.

BACKGROUND: African trypanosomiases are vector-borne diseases that affect humans and livestock in sub-Saharan Africa. Although data have been collected on tsetse fauna as well as trypanosome infections in tsetse flies and mammals in foci of sleeping sickness in Chad, the situation of tsetse fly-transmitted trypanosomes remains unknown in several tsetse-infested areas of Chad. This study was designed to fill this epidemiological knowledge gap by determining the tsetse fauna as well as the trypanosomes infecting tsetse flies in the area of Lake Iro in southeastern Chad. METHODS: Tsetse flies were trapped along the Salamat River using biconical traps. The proboscis and tsetse body were removed from each fly. DNA was extracted from the proboscis using proteinase K and phosphate buffer and from the tsetse body using Chelex 5%. Tsetse flies were identified by amplifying and sequencing the cytochrome c oxydase I gene of each tsetse fly. Trypanosome species were detected by amplifying and sequencing the internal transcribed spacer 1 of infecting trypanosomes. RESULTS: A total of 617 tsetse flies were trapped; the apparent density of flies per trap per day was 2. 6. Of the trapped flies, 359 were randomly selected for the molecular identification and for the detection of infecting trypanosomes. Glossina morsitans submorsitans (96.1%) was the dominant tsetse fly species followed by G. fuscipes fuscipes (3.1%) and G. tachinoides (0.8%). Four trypanosome species, including Trypanosoma vivax, T. simiae, T. godfreyi and T. congolense savannah, were detected. Both single infection (56.7%) and mixed infections of trypanosomes (4.6%) were detected in G. m. submorsitans. The single infection included T. simiae (20.5%), T. congolense savannah (16.43%), T. vivax (11.7%) and T. godfreyi (9.8%). The trypanosome infection rate was 61.4% in G. m. submorsitans, 72.7% in G. f. fuscipes and 66.6% in G. tachinoides. Trypanosome infections were more prevalent in tsetse bodies (40.6%) than in the proboscis (16.3%). CONCLUSION: This study revealed the presence of different tsetse species and a diversity of trypanosomes pathogenic to livestock in the area of Lake Iro. The results highlight the risks and constraints that animal African trypanosomiasis pose to livestock breeding and the importance of assessing trypanosome infections in livestock in this area.

Declining atmospheric deposition of heavy metals over the last three decades is reflected in soil and foliage of 97 beech (Fagus sylvatica) stands in the Vienna Woods.

Rigorous studies on long-term changes of heavy metal distribution in forest soils since the implementation of emission controls are rare. Hence, we resampled 97 old-growth beech stands in the Vienna Woods. This study exploits an extensive data set of soil (infiltration zone of stemflow and between trees area) and foliar chemistry from three decades ago. It was hypothesized that declining deposition of heavy metals is reflected in soil and foliar total contents of Pb, Cu, Zn, Ni, Mn and Fe. Mean soil contents of Pb in the stemflow area declined at the highest rate from 223 to 50 mg kg-1 within the last three decades. Soil contents of Pb and Ni decreased significantly both in the stemflow area and the between trees area down to 80-90 cm soil depth from 1984 to 2012. Top soil (0-5 cm) accumulation and simultaneous loss in the lower soil over time for the plant micro nutrients Cu and Zn are suggested to be caused by plant uptake from deep horizons. Reduced soil leaching, due to a mean soil pH (H2O) increase from 4.3 to 4.9, and increased plant cycling are put forward to explain the significant increase of total Mn contents in the infiltration zone of beech stemflow. Top soil Pb contents in the stemflow area presently exceed the critical value at which toxicity symptoms may occur at numerous sites. Mean foliar contents of all six studied heavy metals decreased within the last three decades, but plant supply with the micro nutrients Cu, Zn, Mn and Fe is still in the optimum range for beech trees. It is concluded that heavy metal pollution is not critical for the studied beech stands any longer.

A slight recovery of soils from Acid Rain over the last three decades is not reflected in the macro nutrition of beech (Fagus sylvatica) at 97 forest stands of the Vienna Woods.

Rigorous studies of recovery from soil acidification are rare. Hence, we resampled 97 old-growth beech stands in the Vienna Woods. This study exploits an extensive data set of soil (infiltration zone of stemflow and between trees area at different soil depths) and foliar chemistry from three decades ago. It was hypothesized that declining acidic deposition is reflected in soil and foliar chemistry. Top soil pH within the stemflow area increased significantly by 0.6 units in both H2O and KCl extracts from 1984 to 2012. Exchangeable Ca and Mg increased markedly in the stemflow area and to a lower extent in the top soil of the between trees area. Trends of declining base cations in the lower top soil were probably caused by mobilization of organic S and associated leaching with high amounts of sulfate. Contents of C, N and S decreased markedly in the stemflow area from 1984 to 2012, suggesting that mineralization rates of organic matter increased due to more favorable soil conditions. It is concluded that the top soil will continue to recover from acidic deposition. However, in the between trees areas and especially in deeper soil horizons recovery may be highly delayed. The beech trees of the Vienna Woods showed no sign of recovery from acidification although S deposition levels decreased. Release of historic S even increased foliar S contents. Base cation levels in the foliage declined but are still adequate for beech trees. Increasing N/nutrient ratios over time were considered not the result of marginally higher N foliar contents in 2012 but of diminishing nutrient uptake due to the decrease in ion concentration in soil solution. The mean foliar N/P ratio already increased to the alarming value of 31. Further nutritional imbalances will predispose trees to vitality loss.

Does mixing of beech (Fagus sylvatica) and spruce (Picea abies) litter hasten decomposition?

BACKGROUND AND AIMS: It is of practical relevance to know how much beech must be admixed to pure spruce stands in order to increase litter decomposition and associated nutrient cycling, since the formation of thick organic layers is commonly ascribed to the recalcitrance of spruce needles. We addressed the impact of tree species mixture within forest stands and within litter on mass loss and nutritional release from litter. METHODS: Litter decomposition was measured in three adjacent stands of pure spruce (Picea abies), mixed beech-spruce and pure beech (Fagus sylvatica) on a nutrient-rich site and a nutrient-poor site over a 2-year period using litterbags which were filled with five different mixtures of beech and spruce litter. RESULTS: Mass loss of beech litter was not higher than mass loss of spruce litter. Decay was primarily affected by tree species composition of the incubation stand and was faster in (mixed) beech forests stands than in spruce forests, while the influence of litter species and their mixtures on decay rates was small. Net transfers of nutrients between the two litter species (direct effects) in the mixed bags were minimal, since initial beech and spruce litter did not have different litter quality. However, in a few cases indirect effects (e.g., changing decomposer abundance and activity) caused non-additive patterns for the totals within the mixed bags, hastening decomposition within the first year. CONCLUSIONS: Greater accumulation of litter in spruce compared to beech stands is not a consequence of the inherent recalcitrance of needles. Adverse environmental conditions in spruce stands retard decomposition. Indirect effects on decomposition caused by stand mixture are not mimicked by litter mixtures within mesh bags.

Greater accumulation of litter in spruce (Picea abies) compared to beech (Fagus sylvatica) stands is not a consequence of the inherent recalcitrance of needles.

BACKGROUND AND AIMS: Replacement of beech by spruce is associated with changes in soil acidity, soil structure and humus form, which are commonly ascribed to the recalcitrance of spruce needles. It is of practical relevance to know how much beech must be admixed to pure spruce stands in order to increase litter decomposition and associated nutrient cycling. We addressed the impact of tree species mixture within forest stands and within litter on mass loss and nutritional release from litter. METHODS: Litter decomposition was measured in three adjacent stands of pure spruce (Picea abies), mixed beech-spruce and pure beech (Fagus sylvatica) on three nutrient-rich sites and three nutrient-poor sites over a three-year period using the litterbag method (single species and mixed species bags). RESULTS: Mass loss of beech litter was not higher than mass loss of spruce litter. Mass loss and nutrient release were not affected by litter mixing. Litter decay indicated non-additive patterns, since similar remaining masses under pure beech (47%) and mixed beech-spruce (48%) were significantly lower than under pure spruce stands (67%). Release of the main components of the organic substance (Corg, Ntot, P, S, lignin) and associated K were related to mass loss, while release of other nutrients was not related to mass loss. CONCLUSIONS: In contradiction to the widely held assumption of slow decomposition of spruce needles, we conclude that accumulation of litter in spruce stands is not caused by recalcitrance of spruce needles to decay; rather adverse environmental conditions in spruce stands retard decomposition. Mixed beech-spruce stands appear to be as effective as pure beech stands in counteracting these adverse conditions.