Evolution of cassava brown streak disease-associated viruses.

Cassava brown streak disease (CBSD) has occurred in the Indian Ocean coastal lowlands and some areas of Malawi in East Africa for decades, and makes the storage roots of cassava unsuitable for consumption. CBSD is associated with Cassava brown streak virus (CBSV) and the recently described Ugandan cassava brown streak virus (UCBSV) [picorna-like (+)ssRNA viruses; genus Ipomovirus; family Potyviridae]. This study reports the first comprehensive analysis on how evolution is shaping the populations of CBSV and UCBSV. The complete genomes of CBSV and UCBSV (four and eight isolates, respectively) were 69.0-70.3 and 73.6-74.4% identical at the nucleotide and polyprotein amino acid sequence levels, respectively. They contained predictable sites of homologous recombination, mostly in the 3'-proximal part (NIb-HAM1h-CP-3'-UTR) of the genome, but no evidence of recombination between the two viruses was found. The CP-encoding sequences of 22 and 45 isolates of CBSV and UCBSV analysed, respectively, were mainly under purifying selection; however, several sites in the central part of CBSV CP were subjected to positive selection. HAM1h (putative nucleoside triphosphate pyrophosphatase) was the least similar protein between CBSV and UCBSV (aa identity approx. 55%). Both termini of HAM1h contained sites under positive selection in UCBSV. The data imply an on-going but somewhat different evolution of CBSV and UCBSV, which is congruent with the recent widespread outbreak of UCBSV in cassava crops in the highland areas (>1000 m above sea level) of East Africa where CBSD has not caused significant problems in the past.

Simultaneous virus-specific detection of the two cassava brown streak-associated viruses by RT-PCR reveals wide distribution in East Africa, mixed infections, and infections in Manihot glaziovii.

The expanding cassava brown streak disease (CBSD) epidemic in East Africa is caused by two ipomoviruses (genus Ipomovirus; Potyviridae), namely, Cassava brown streak virus (CBSV), and Ugandan cassava brown streak virus (UCBSV) that was described recently. A reverse transcription polymerase chain reaction (RT-PCR) based diagnostic method was developed in this study for simultaneous virus-specific detection of the two viruses. Results showed that CBSV and UCBSV are distributed widely in the highlands (> 1000 m above the sea level) of the Lake Victoria zone in Uganda and Tanzania and also in the Indian Ocean costal lowlands of Tanzania. Isolates of UCBSV from the Lake Victoria zone were placed to two phylogenetic clusters in accordance with their origin in Uganda or Tanzania, respectively. Mixed infections with CBSV and UCBSV were detected in many cassava plants in the areas surveyed. CBSV was also detected in the perennial species Manihot glaziovii (DNA-barcoded in this study) in Tanzania, which revealed the first virus reservoir other than cassava. The method for detection of CBSV and UCBSV described in this study has important applications for plant quarantine, resistance breeding of cassava, and studies on epidemiology and control of CBSD in East Africa.

First Report of Sweetpotato symptomless virus 1 and Sweetpotato virus A in Sweetpotatoes in Tanzania.

Sweetpotato (Ipomea batatas L.) is grown widely from tropical to temperate regions and is an important food security crop in tropical countries. In Africa, sweetpotato is infected by RNA viruses of many taxa (4), but DNA viruses, such as the genus Begomovirus (family Geminiviridae), infecting sweetpotatoes in the Americas have been reported only in Kenya (3). A caulimo-like DNA virus (family Caulimoviridae) has been detected in sweetpotatoes in Uganda (1). Recently, two novel badnaviruses (genus Badnavirus, family Caulimoviridae) and a new mastrevirus (genus Mastrevirus, family Geminiviridae) were discovered in a local sweetpotato cultivar maintained in a germplasm collection in Peru (2) but were not reported elsewhere. This study examined the possible existence of these novel viruses in landrace sweetpotato varieties grown in Tanzania. Nine landrace sweetpotato varieties and one introduced cultivar (NIS 91 from the International Potato Centre, Peru) were sampled from six regions of Tanzania. DNA was extracted (2) and amplified by PCR using primers (MastvKF: 5'-GACAGACCCCTAGGGTGA-3'; MastvsR 5'-ACTGCATATAGTACATGCCACA-3') designed in this study to amplify partial, putative movement and coat protein gene sequences of Sweetpotato symptomless virus 1 (SPSMV-1) (GenBank Accession No. FJ560945) (2). Products of the expected size were detected in seven samples (varieties Ex-London, Ex-Lyawaya, Gairo, Hombolo, Kagole white, Mbeya, and Shangazi) representing four regions surveyed (Dodoma, Mbeya, Morogoro, and Kagera). PCR products from five samples were sequenced (396 nt; GenBank Accession Nos. HQ316938 to HQ316942) and found to be identical to each other and the isolate described originally in Peru (2). Amplification with primers (BadnaBKF: 5'-CAAATTAGGAGGCAGATAAATG-3'; BadnaBsR: 5'-GGTCTTCTTATGTTCCACCTT-3') designed in this study according to the sequence of Sweetpotato virus B (SPBV-B) (GenBank Accession No.FJ560944) resulted in products of the expected size in three samples (varieties Ex-Lyawaya, Gairo, and Hombolo collected in Mbeya, Morogoro, and Dodoma, respectively) that were positive also for SPSMV-1. Sequences of the products (787 nt; HQ316935 to HQ316937) were nearly identical (99.4%). They were 96.8 to 96.9% similar to a region (nts 830-1616) of Sweetpotato virus A (SPBV-A; FJ560943) (2), whereas they were only 83.2 to 83.6 % similar to the corresponding region (1,486 to 2,272 nt) of SPBV-B (FJ560944) (2). No virus was detected in cv. NIS 91. All plants sampled exhibited mild mottling or mosaic symptoms, but a contribution to the symptoms by other untested viruses cannot be excluded because few of the large number of sweetpotato viruses have been studied in Africa (4). To our knowledge, this is the first report of SPSMV-1 and SPBV-A outside South America and in sweetpotatoes grown in the field. The results show that the two viruses are distributed widely in local sweetpotato varieties in Tanzania, which suggests that they may be found in other sweetpotato-growing areas where they have not been studied. While the yield losses caused by SPSMV-1 and SPBV-A remains to be studied, the data from this study are of practical importance in terms of regional and international exchange of sweetpotato germplasm. References: (1) V. Aritua et al. Plant Pathol. 56:324, 2007. (2) J. F. Kreuze et al. Virology 388:1, 2009. (3) T. Paprotka et al. Virus Res. 149:224, 2010. (4) F. Tairo et al. Mol. Plant Pathol. 6:199, 2005.

Genetically distinct strains of Cassava brown streak virus in the Lake Victoria basin and the Indian Ocean coastal area of East Africa.

Six isolates of Cassava brown streak virus (CBSV, genus Ipomovirus; Potyviridae) from the Lake Victoria basin in Uganda and Tanzania were characterized. Virus particles were 650 nm long. The complete coat protein (CP)-encoding sequences (1,101 nucleotides, nt) were 90.7-99.5 and 93.7-99.5% identical at the nt and amino acid (aa) levels, respectively. The 3' untranslated region was 225, 226 or 227 nt long. These eight isolates were only 75.8-77.5% (nt) and 87.0-89.9% (aa) identical when compared to the partial CP sequences (714 nt) of six CBSV isolates characterized previously from the costal lowlands of Tanzania and Mozambique. Hence, two genetically different and geographically separated populations of CSBV exist in East Africa.

Identification of Candida strains isolated from Tanzanian pregnant women with vaginal candidiasis.

OBJECTIVE: To identify Candida strains isolated from Tanzanian women (13 to 45 years) with vaginal candidiasis. DESIGN: A cross-sectional study. SETTING: Antenatal clinic in Ilala district hospital in Dar es Salaam, Tanzania from March 1998 to December 2000. RESULTS: The identities of the 272 isolates tested with API Candida were: Candida albicans 180(66.2%), Candida tropicalis 13(4.7%), Candida glabrata 20(7.35%), Candida famata 6(2.2%), Candida parapsilosis 6 (2.2%) and Candida lusitaniae one (0.37%). API Candida could not speciate 43 (15.8%) isolates of these; two (0.7%) fell between C. albicans and C. tropicalis, 17(6.25%) C. lusitaniael, C. guilliermondii/C. famata, 14(5.15%) C. krusei, C. inconspicua, and C. norvegensis and nine (3.3%) either C. parapsilosis, C. krusei, C. incospicua or Geotrichum spp. Four (1.5%) isolates had an assimilation pattern of Trichosporo spp, but were all germ tube positive and had morphological features on cornemeal agar that were consistent with C. albicans. API 20C AUX was used for testing 29 isolates and results showed: 11/29 (37.9%) C. albicans, 1/29 (3.4%) C. tropicalis, 4/29 (13.8%) C. glabrata, 1/29 (3.4%) C. parapsilosis, 1/29 (3.4%) C. famata, 1/29 (3.4%) C. lusitaniae, 1/29 (3.4%) C. colliculosa/C. magnoliae, 5/29(17.2%) C. albicans/ C. tropicals 2/29 (6.8%) C. norvegensis/C. parapsilosis, and 2/29(6.8%) C. kruseil/C. inconspicua. Results of 20 isolates identified by Randomly Amplified Polymorphic DNA (RAPID) technique showed a 95% agreement with API Candida and a 100% agreement with API 20C AUX. CONCLUSION: Although most (66.3%) of the species isolated from Dar es Salaam women with vaginal candidiasis were C. albicans, a considerable percentage (33.7%) were non-albicans, mainly C. glabrata, C. krusei and C. tropicalis. The high prevalence of non-albicans Candida spp observed in this population may have therapeutic implications.

Coat protein sequence analysis reveals occurrence of new strains of Sweet potato feathery mottle virus in Uganda and Tanzania.

The 3'-proximal part (1.8 kb) of the Sweet potato feathery mottle virus (SPFMV) genome was studied in four SPFMV isolates collected from farmers' fields in western Uganda (SPFMV-Bny), eastern Uganda (SPFMV-Sor) and Bagamoyo district, Tanzania (SPFMV-TZ1 and SPFMV-TZ2). Unlike the other three SPFMV isolates, SPFMV-Sor was not detected with the polyclonal antisera to SPFMV. It showed moderately high coat protein (CP) nucleotide (93.3-96.7%) and amino acid (93.6-96.8%) sequence identity to the isolates of the SPFMV strain group C. In contrast, identities (78.1-80.1%, and 79.9-83.1%) to isolates of the SPFMV strain groups O, RC, and the East African (EA) strain group were low. Similar to some isolates (SPFMV-CH2 and SPFMV-6) of strain group C, but different from other SPFMV isolates, SPFMV-Sor contained a deletion of 6 nucleotides in the CP-encoding region (CP amino acid positions 62-63). Phylogenetic analysis of the CP sequences indicated that SPFMV-Sor belongs to the SPFMV strain group C that has not been reported from Africa. Sequence data were obtained for the first time from Tanzanian SPFMV isolates in this study, and phylogenetic analysis indicated that they belong to the strain group EA, which is unique to East Africa.