Expansion of the Cassava Brown Streak Disease Epidemic in Eastern Democratic Republic of Congo.

Cassava plays a key role in ensuring food security and generating income for smallholder farmers throughout Central Africa, particularly in the Democratic Republic of Congo (DRC). This status is threatened, however, by cassava brown streak disease (CBSD), which has expanded its incidence and range in eastern DRC. The study described here comprises the first extensive assessment of temporal change in the occurrence of CBSD and its causal viruses in DRC, based on surveys conducted during 2016 and 2018. Cassava fields were inspected in Ituri, Nord-Kivu, Sud-Kivu, Tanganyika, and Haut-Katanga provinces within eastern DRC to record foliar incidence and severity of CBSD. Leaf samples were collected for virus detection and species-level identification. New occurrences of CBSD, confirmed by virus diagnostic tests, were recorded in two provinces (Haut-Katanga and Sud-Kivu) and nine previously unaffected territories, covering an area of >62,000 km2, and at up to 900 km from locations of previously published reports of CBSD in DRC. Overall, average CBSD incidence within fields was 13.2% in 2016 and 16.1% in 2018. In the new spread zone of Haut-Katanga, incidence increased from 1.7 to 15.9%. CBSD is now present in provinces covering 321,000 km2, which is approximately 14% of the total area of DRC. This represents a major expansion of the CBSD epidemic, which was only recorded from one province (Nord-Kivu) in 2012. Both Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus were detected in Ituri, Nord-Kivu, and Sud-Kivu, but only CBSV was detected in Haut-Katanga. Overall, these results confirm the increasing threat that CBSD poses to cassava production in DRC and describe an important expansion in the African pandemic of CBSD.

Spatial Analysis of Temporal Changes in the Pandemic of Severe Cassava Mosaic Disease in Northwestern Tanzania.

To improve understanding of the dynamics of the cassava mosaic disease (CMD) pandemic front, geospatial approaches were applied to the analysis of 3 years' data obtained from a 2-by-2° (approximately 222-by-222 km) area of northwestern Tanzania. In total, 80 farmers' fields were assessed in each of 2009, 2010, and 2011, with 20 evenly distributed fields per 1-by-1° quadrant. CMD-associated variables (CMD incidence, CMD severity, vector-borne CMD infection, and vector abundance) increased in magnitude from 2009 to 2010 but showed little change from 2010 to 2011. Increases occurred primarily in the two westernmost quadrants of the study area. A pandemic "front" was defined by determining the values of CMD incidence and whitefly abundance where predicted disease gradients were greatest. The pandemic-associated virus (East African cassava mosaic virus-Uganda) and vector genotype (Bemisia tabaci sub-Saharan Africa 1-subgroup 1) were both present within the area bounded by the CMD incidence front but both also occurred ahead of the front. The average speed and direction of movement of the CMD incidence front (22.9 km/year; southeast) and whitefly abundance front (46.6 km/year; southeast) were calculated, and production losses due to CMD were estimated to range from US$4.3 million to 12.2 million.

A Risk Assessment Framework for Seed Degeneration: Informing an Integrated Seed Health Strategy for Vegetatively Propagated Crops.

Pathogen buildup in vegetative planting material, termed seed degeneration, is a major problem in many low-income countries. When smallholder farmers use seed produced on-farm or acquired outside certified programs, it is often infected. We introduce a risk assessment framework for seed degeneration, evaluating the relative performance of individual and combined components of an integrated seed health strategy. The frequency distribution of management performance outcomes was evaluated for models incorporating biological and environmental heterogeneity, with the following results. (1) On-farm seed selection can perform as well as certified seed, if the rate of success in selecting healthy plants for seed production is high; (2) when choosing among within-season management strategies, external inoculum can determine the relative usefulness of 'incidence-altering management' (affecting the proportion of diseased plants/seeds) and 'rate-altering management' (affecting the rate of disease transmission in the field); (3) under severe disease scenarios, where it is difficult to implement management components at high levels of effectiveness, combining management components can be synergistic and keep seed degeneration below a threshold; (4) combining management components can also close the yield gap between average and worst-case scenarios. We also illustrate the potential for expert elicitation to provide parameter estimates when empirical data are unavailable. [Formula: see text] Copyright © 2017 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Analysis of Interactions Between Cassava Brown Streak Disease Symptom Types Facilitates the Determination of Varietal Responses and Yield Losses.

Cassava brown streak disease (CBSD), caused by cassava brown streak viruses, is recognized as one of the most important plant disease threats to African food security. This study describes the incidence and severity of the different symptom types caused by CBSD, derived from extensive surveys in the country most severely affected by the disease: Tanzania. Total plant incidence and mean root severity of CBSD, recorded from 341 farmers' fields, were both greater in the Coast Zone (49.5% and 3.05), than in the Lake Zone (32.7% and 2.57). Overall, the differing incidences recorded declined in the following order: total plant incidence (39.1%), plant shoot incidence (33.4%), plant root incidence (19.3%), root incidence (10.5%), and unusable root incidence (5.4%). The much lower-than-anticipated loss due to the root necrosis that is characteristic of CBSD was offset by large reductions of root number in plants expressing foliar symptoms of CBSD (15.7% in the Coast Zone and 5.5% in the Lake Zone). These data suggest that the effects of CBSD on the growth of affected plants are greater than those due to root spoilage. Based on these two factors, annual losses due to CBSD in the parts of Tanzania surveyed were estimated at >860,000 t, equivalent to more than U.S.$51 million. A novel approach to using farm-derived data on the responses of the most frequently cultivated varieties to CBSD infection allowed the grouping of the varieties into four categories, based on their relative resistance or tolerance to infection. This tool should be of value to breeders in identifying and selecting for sources of resistance or tolerance in both local and exotic germplasm, and should ultimately contribute to enhancing the management of one of Africa's most damaging crop diseases.

The Dynamics and Environmental Influence on Interactions Between Cassava Brown Streak Disease and the Whitefly, Bemisia tabaci.

Cassava brown streak disease (CBSD) is currently the most significant virus disease phenomenon affecting African agriculture. In this study, we report results from the most extensive set of field data so far presented for CBSD in Africa. From assessments of 515 farmers' plantings of cassava, incidence in the Coastal Zone of Tanzania (46.5% of plants; 87% of fields affected) was higher than in the Lake Zone (22%; 34%), but incidences for both zones were greater than previous published records. The whitefly vector, Bemisia tabaci, was more abundant in the Lake Zone than the Coastal Zone, the reverse of the situation reported previously, and increased B. tabaci abundance is driving CBSD spread in the Lake Zone. The altitudinal "ceiling" previously thought to restrict the occurrence of CBSD to regions <1,000 masl has been broken as a consequence of the greatly increased abundance of B. tabaci in mid-altitude areas. Among environmental variables analyzed, minimum temperature was the strongest determinant of CBSD incidence. B. tabaci in the Coastal and Lake Zones responded differently to environmental variables examined, highlighting the biological differences between B. tabaci genotypes occurring in these regions and the superior adaptation of B. tabaci in the Great Lakes region both to cassava and low temperature conditions. Regression analyses using multi-country data sets could be used to determine the potential environmental limits of CBSD. Approaches such as this offer potential for use in the development of predictive models for CBSD, which could strengthen country- and continent-level CBSD pandemic mitigation strategies.

Impact of antibiotic treatment for pulmonary exacerbations on bacterial diversity in cystic fibrosis.

BACKGROUND: A diverse array of bacterial species is present in the CF airways, in addition to those recognised as clinically important. Here, we investigated the relative impact of antibiotics, used predominantly to target Pseudomonas aeruginosa during acute exacerbations, on other non-pseudomonal species. METHODS: The relative abundance of viable P. aeruginosa and non-pseudomonal species was determined in sputa from 12 adult CF subjects 21, 14, and 7 days prior to antibiotics, day 3 of treatment, the final day of treatment, and 10-14 days afterwards, by T-RFLP profiling. RESULTS: Overall, relative P. aeruginosa abundance increased during antibiotic therapy compared to other bacterial species; mean abundance pre-antibiotic 51.0±36.0% increasing to 71.3±30.4% during antibiotic (ANOVA: F(1,54)=5.16; P<0.027). Further, the number of non-pseudomonal species detected fell; pre-antibiotic 6.0±3.3 decreasing to 3.7±3.3 during treatment (ANOVA: F(1,66)=5.11; P<0.027). CONCLUSIONS: Antibiotic treatment directed at P. aeruginosa has an additional significant impact on non-pseudomonal, co-colonising species.

Multiplex RT-PCR assays for the simultaneous detection of both RNA and DNA viruses infecting cassava and the common occurrence of mixed infections by two cassava brown streak viruses in East Africa.

Uniplex and multiplex reverse transcription-polymerase chain reaction (RT-PCR) protocols were developed for the detection of cassava brown streak viruses (CBSVs) in single and mixed infections with cassava mosaic begomoviruses (CMBs) in a tropical crop plant, cassava (Manihot esculenta). CMBs contain ssDNA as their genome (genus Begomovirus, family Geminiviridae) while CBSVs are made up of positive sense ssRNA (genus Ipomovirus, family Potyviridae), and they cause the economically important cassava mosaic and cassava brown streak diseases, respectively, in sub-Saharan Africa. Diagnostic methodologies have long been available for CMBs but they are limited for CBSVs especially in mixed infections. In this study, the two CBSVs, Cassava brown streak virus (CBSV) and Cassava brown streak Uganda virus (CBSUV) occurring singly or in mixed infection with CMBs, African cassava mosaic virus and East African cassava mosaic virus were detected in a single RT-PCR using both previously described and newly designed virus-specific primers. These protocols were highly efficient for detecting CBSVs compared to the existing methods and have great potential to minimize sample handling and contamination. As well as improving the diagnosis of cassava viruses, the development of multiplex RT-PCR protocols have revealed the common occurrence of mixed infections by CBSV and CBSUV in cassava fields of Tanzania and Kenya, which was contrary to the common belief until recently that these two viruses have existed separately. These protocols have implications for diagnosis and epidemiological studies on cassava virus diseases in Eastern Africa.

Widespread occurrence and diversity of Cassava brown streak virus (Potyviridae: Ipomovirus) in Tanzania.

Cassava brown streak disease (CBSD) has been a problem in Tanzania since 1936. Existing literature indicated limited distribution of the disease to low altitudes, usually <100 m above sea level, but the current geographical distribution of the disease was not known. Whether a single or many strains for the virus exist in Tanzania had not been reported to date. In this study, CBSD was recorded from sea level to ≈1,800 m above sea level. In total, 2,730 cassava plants were assessed for CBSD leaf symptoms in 91 fields and root symptoms were assessed at 81 sites. A sample was taken from each site for laboratory screening for Cassava brown streak virus (CBSV). CBSD mean foliar and root incidences were 38 and 36%, respectively. Reverse-transcription polymerase chain reaction of a partial 3'-terminal coat protein (CP) region of CBSV indicated the presence of CBSV in 67 of the 91 (73%) samples. Forty-three amplicons were sequenced, and phylogenetic comparisons with nucleotide sequences from GenBank (National Center for Biotechnology Information database) suggested that one major clade of CBSV primarily exists in Tanzania. However, there was nucleotide sequence divergence of up to 19% among the 42 isolates. In all, 42 of the 43 sequences had 80 to 100% nucleotide identity with 6 previously reported CP-CBSV sequences (from Mozambique and Tanzania). In total, 13 of 42 isolates had <80% nucleotide identities with three previously reported Ugandan CBSV sequences. One isolate, FJ687177, shared <78% sequence identity with the other Tanzanian sequences but was closely related (93%) to Ugandan isolates. It is likely that isolate FJ687177 may belong to a less widely distributed recently described species (clade 2) of CBSV, named Ugandan cassava brown streak virus (UCBSV).

Comparing the regional epidemiology of the cassava mosaic and cassava brown streak virus pandemics in Africa.

The rapid geographical expansion of the cassava mosaic disease (CMD) pandemic, caused by cassava mosaic geminiviruses, has devastated cassava crops in 12 countries of East and Central Africa since the late 1980s. Region-level surveys have revealed a continuing pattern of annual spread westward and southward along a contiguous 'front'. More recently, outbreaks of cassava brown streak disease (CBSD) were reported from Uganda and other parts of East Africa that had been hitherto unaffected by the disease. Recent survey data reveal several significant contrasts between the regional epidemiology of these two pandemics: (i) severe CMD radiates out from an initial centre of origin, whilst CBSD seems to be spreading from independent 'hot-spots'; (ii) the severe CMD pandemic has arisen from recombination and synergy between virus species, whilst the CBSD pandemic seems to be a 'new encounter' situation between host and pathogen; (iii) CMD pandemic spread has been tightly linked with the appearance of super-abundant Bemisia tabaci whitefly vector populations, in contrast to CBSD, where outbreaks have occurred 3-12 years after whitefly population increases; (iv) the CMGs causing CMD are transmitted in a persistent manner, whilst the two cassava brown streak viruses appear to be semi-persistently transmitted; and (v) different patterns of symptom expression mean that phytosanitary measures could be implemented easily for CMD but have limited effectiveness, whereas similar measures are difficult to apply for CBSD but are potentially very effective. An important similarity between the pandemics is that the viruses occurring in pandemic-affected areas are also found elsewhere, indicating that contrary to earlier published conclusions, the viruses per se are unlikely to be the key factors driving the two pandemics. A diagrammatic representation illustrates the temporal relationship between B. tabaci abundance and changing incidences of both CMD and CBSD in the Great Lakes region. This emphasizes the pivotal role played by the vector in both pandemics and the urgent need to identify effective and sustainable strategies for controlling whiteflies on cassava.

Optimization of diagnostic RT-PCR protocols and sampling procedures for the reliable and cost-effective detection of Cassava brown streak virus.

Sampling procedures and diagnostic protocols were optimized for accurate diagnosis of Cassava brown streak virus (CBSV) (genus Ipomovirus, family Potyviridae). A cetyl trimethyl ammonium bromide (CTAB) method was optimized for sample preparation from infected cassava plants and compared with the RNeasy plant mini kit (Qiagen) for sensitivity, reproducibility and costs. CBSV was detectable readily in total RNAs extracted using either method. The major difference between the two methods was in the cost of consumables, with the CTAB 10x cheaper (0.53 pounds sterling=US$0.80 per sample) than the RNeasy method (5.91 pounds sterling=US$8.86 per sample). A two-step RT-PCR (1.34 pounds sterling=US$2.01 per sample), although less sensitive, was at least 3-times cheaper than a one-step RT-PCR (4.48 pounds sterling=US$6.72). The two RT-PCR tests revealed consistently the presence of CBSV both in symptomatic and asymptomatic leaves and indicated that asymptomatic leaves can be used reliably for virus diagnosis. Depending on the accuracy required, sampling 100-400 plants per field is an appropriate recommendation for CBSD diagnosis, giving a 99.9% probability of detecting a disease incidence of 6.7-1.7%, respectively. CBSV was detected at 10(-4)-fold dilutions in composite sampling, indicating that the most efficient way to index many samples for CBSV will be to screen pooled samples. The diagnostic protocols described below are reliable and the most cost-effective methods available currently for detecting CBSV.

Molecular biodiversity of cassava begomoviruses in Tanzania: evolution of cassava geminiviruses in Africa and evidence for East Africa being a center of diversity of cassava geminiviruses.

Cassava is infected by numerous geminiviruses in Africa and India that cause devastating losses to poor farmers. We here describe the molecular diversity of seven representative cassava mosaic geminiviruses (CMGs) infecting cassava from multiple locations in Tanzania. We report for the first time the presence of two isolates in East Africa: (EACMCV-[TZ1] and EACMCV-[TZ7]) of the species East African cassava mosaic Cameroon virus, originally described in West Africa. The complete nucleotide sequence of EACMCV-[TZ1] DNA-A and DNA-B components shared a high overall sequence identity to EACMCV-[CM] components (92% and 84%). The EACMCV-[TZ1] and -[TZ7] genomic components have recombinations in the same genome regions reported in EACMCV-[CM], but they also have additional recombinations in both components. Evidence from sequence analysis suggests that the two strains have the same ancient origin and are not recent introductions. EACMCV-[TZ1] occurred widely in the southern part of the country. Four other CMG isolates were identified: two were close to the EACMV-Kenya strain (named EACMV-[KE/TZT] and EACMV-[KE/TZM] with 96% sequence identity); one isolate, TZ10, had 98% homology to EACMV-UG2Svr and was named EACMV-UG2 [TZ10]; and finally one isolate was 95% identical to EACMV-[TZ] and named EACMV-[TZ/YV]. One isolate of African cassava mosaic virus with 97% sequence identity with other isolates of ACMV was named ACMV-[TZ]. It represents the first ACMV isolate from Tanzania to be sequenced. The molecular variability of CMGs was also evaluated using partial B component nucleotide sequences of 13 EACMV isolates from Tanzania. Using the sequences of all CMGs currently available, we have shown the presence of a number of putative recombination fragments that are more prominent in all components of EACMV than in ACMV. This new knowledge about the molecular CMG diversity in East Africa, and in Tanzania in particular, has led us to hypothesize about the probable importance of this part of Africa as a source of diversity and evolutionary change both during the early stages of the relationship between CMGs and cassava and in more recent times. The existence of multiple CMG isolates with high DNA genome diversity in Tanzania and the molecular forces behind this diversity pose a threat to cassava production throughout the African continent.

Methods of surveying the incidence and severity of cassava mosaic disease and whitefly vector populations on cassava in Africa: a review.

Field surveys in many cassava growing areas of Africa have assessed the incidence and severity of cassava mosaic disease (CMD), populations of the whitefly vector (Bemisia tabaci), and the distribution of cassava mosaic begomoviruses (CMBs). The methods employed differ greatly between countries and attempts at standardization were made in recent CMD surveys in East and Central Africa, notably in the systemwide Whitefly IPM Project, which provides a paradigm for future work on CMBs and whiteflies on cassava in Africa and also elsewhere. However, there is a need for greater standardization so as to assess the continued expansion of the current CMD pandemic in eastern Africa. Standardized methods will facilitate the collection of reliable data, which can be used to predict future disease spread, develop appropriate management strategies and compare disease development between seasons and locations. In this review, the methods used and the problems encountered during such surveys are discussed and recommendations made on future procedure.

Cassava mosaic geminiviruses in Africa.

Cassava mosaic disease (CMD) caused by cassava mosaic geminiviruses (CMGs) (Geminiviridae:Begomovirus) is undoubtedly the most important constraint to the production of cassava in Africa at the outset of the 21st century. Although the disease was recorded for the first time in the latter part of the 19th century, for much of the intervening period it has been relatively benign in most of the areas where it occurs and has generally been considered to be of minor economic significance. Towards the end of the 20th century, however, the inherent dynamism of the causal viruses was demonstrated, as a recombinant hybrid of the two principal species was identified, initially from Uganda, and shown to be associated with an unusually severe and rapidly spreading epidemic of CMD. Subsequent spread throughout East and Central Africa, the consequent devastation of production of the cassava crop, a key staple in much of this region, and the observation of similar recombination events elsewhere, has once again demonstrated the inherent danger posed to man by the capacity of these viruses to adapt to their environment and optimally exploit their relationships with the whitefly vector, plant host and human cultivator. In this review of cassava mosaic geminiviruses in Africa, we examine each of these relationships, and highlight the ways in which the CMGs have exploited them to their own advantage.

A distinct Bemisia tabaci (Gennadius) (Hemiptera: Sternorrhyncha: Aleyrodidae) genotype cluster is associated with the epidemic of severe cassava mosaic virus disease in Uganda.

During the 1990s, an epidemic of cassava mosaic virus disease caused major losses to cassava production in Uganda. Two factors associated with the epidemic were the occurrence of a novel recombinant begomovirus, EACMV-Ug, and unusually high populations of the whitefly vector, Bemisia tabaci. Here we present molecular evidence for the occurrence of two cassava-colonizing B. tabaci genotype clusters, Ug1 and Ug2, one of which, Ug2, can be consistently associated with the CMD epidemic in Uganda at the time of collection in 1997. By contrast, a second genotype cluster, Ug1, only occurred 'at' or 'ahead of' the epidemic 'front', sometimes in mixtures with Ug2. Comparison of mitochondrial cytochrome oxidase I gene sequences for Ug1 and Ug2 and well-studied B. tabaci reference populations indicated that the two Ugandan populations exhibited approximately 8% divergence, suggesting they represent distinct sub-Saharan African lineages. Neither Ugandan genotype cluster was identified as the widely distributed, polyphagous, and highly fecund B biotype of Old World origin, with which they both diverged by approximately 8%. Within genotype cluster divergence of Ug1 at 0.61 +/- 0.1% was twice that of Ug2 at 0.35 +/- 0.1%. Mismatch analysis suggested that Ug2 has undergone a recent population expansion and may be of nonUgandan origin, whereas Ug1 has diverged more slowly, and is likely to be an indigenous genotype cluster.

A hypothesis: antenatal sensitisation to respiratory syncytial virus in viral bronchiolitis.

AIM: To investigate the possibility of antenatal sensitisation to respiratory syncytial virus (RSV). METHODS: A total of 36 cord blood specimens were obtained from newborn infants; serum IgA was measured to exclude maternal blood contamination. Cord peripheral blood mononuclear cells were separated and cultured in the presence of either uninfected negative control cells or cells containing ultraviolet (UV) inactivated RSV. Proliferation was assessed by tritiated thymidine incorporation. Supernatant cytokine concentrations were measured using ELISA. RESULTS: Significantly higher proliferative response rates to UV inactivated RSV were shown in those infants exposed in utero to the RSV epidemic after 22 weeks gestation. UV inactivated RSV stimulation induced significantly higher interferon gamma production from specimens with a positive proliferative response (sensitised) than from those with a negative response (not sensitised). CONCLUSIONS: Antenatal sensitisation to RSV occurs in one third of infants exposed to an RSV epidemic at the appropriate time of gestation. This sensitisation is associated with increased interferon gamma production, suggesting a type 1 memory response. We hypothesise that priming of fetal T cells to RSV results in a reduced severity of subsequent RSV disease in these individuals and that this will explain much of the clinical diversity of RSV disease.

Cassava mosaic virus disease in East Africa: a dynamic disease in a changing environment.

Cassava mosaic disease (CMD), now known to be caused by cassava mosaic geminiviruses (Family Geminiviridae; Genus Begomovirus), was first reported in East Africa in 1894. Epidemics occurred in Madagascar and Uganda in the 1930s and 1940s, and more localised rapid spread of CMD was observed in parts of coastal Tanzania in the 1930s and coastal Kenya in the 1970s. During the 1990s, a major regional pandemic of an unusually severe form of CMD has expanded to affect parts of at least five countries, causing massive economic losses and destabilising food security. Mechanisms responsible for the development and progress of the pandemic have been described, and comparisons of epidemiological data for varieties grown throughout the period under review suggest that the recent pandemic has been characterised by rapid rates of CMD spread hitherto unknown in East Africa. A key factor in the genesis and spread of the pandemic has been the recombination between two distinct cassava mosaic geminiviruses to produce a novel and more virulent hybrid. Although such events may be common, the known history of CMD in East Africa suggests that the frequency with which they become epidemiologically significant is low. A corollary of this is that resistance, developed originally in Tanzania between 1934 and 1960, and utilized and supplemented at the International Institute of Tropical Agriculture, Nigeria, since 1971, is providing effective CMD control in current pandemic-affected areas of East Africa. Consequently, it is concluded that prospects for managing CMD in the 21st century are good, and that the approach adopted should build on the model of collaborative research and implementation that has been established in tackling the current CMD pandemic.