European origin of Bradyrhizobium populations infecting lupins and serradella in soils of Western Australia and South Africa

We applied a multilocus phylogenetic approach to elucidate the origin of serradella and lupin Bradyrhizobium strains that persist in soils of Western Australia and South Africa. The selected strains belonged to different randomly amplified polymorphic DNA (RAPD)-PCR clusters that were distinct from RAPD clusters of applied inoculant strains. Phylogenetic analyses were performed with nodulation genes (nodA, nodZ, nolL, noeI), housekeeping genes (dnaK, recA, glnII, atpD), and 16S-23S rRNA intergenic transcribed spacer sequences. Housekeeping gene phylogenies revealed that all serradella and Lupinus cosentinii isolates from Western Australia and three of five South African narrow-leaf lupin strains were intermingled with the strains of Bradyrhizobium canariense, forming a well supported branch on each of the trees. All nodA gene sequences of the lupin and serradella bradyrhizobia formed a single branch, referred to as clade II, together with the sequences of other lupin and serradella strains. Similar patterns were detected in nodZ and nolL trees. In contrast, nodA sequences of the strains isolated from native Australian legumes formed either a new branch called clade IV or belonged to clade I or III, whereas their nonsymbiotic genes grouped outside the B. canariense branch. These data suggest that the lupin and serradella strains, including the strains from uncultivated L. cosentinii plants, are descendants of strains that most likely were brought from Europe accidentally with lupin and serradella seeds. The observed dominance of B. canariense strains may be related to this species' adaptation to acid soils common in Western Australia and South Africa and, presumably, to their intrinsic ability to compete for nodulation of lupins and serradella.     

Characterization of Bradyrhizobium strains indigenous to Western Australia and South Africa indicates remarkable genetic diversity and reveals putative new species

Bradyrhizobium are N₂-fixing microsymbionts of legumes with relevant applications in agricultural sustainability, and we investigated the phylogenetic relationships of conserved and symbiotic genes of 21 bradyrhizobial strains. The study included strains from Western Australia (WA), isolated from nodules of Glycine spp. the country is one genetic center for the genus and from nodules of other indigenous legumes grown in WA, and strains isolated from forage Glycine sp. grown in South Africa. The 16S rRNA phylogeny divided the strains in two superclades, of B. japonicum and B. elkanii, but with low discrimination among the species. The multilocus sequence analysis (MLSA) with four protein-coding housekeeping genes (dnaK, glnII, gyrB and recA) pointed out seven groups as putative new species, two within the B. japonicum, and five within the B. elkanii superclades. The remaining eleven strains showed higher similarity with six species, B. lupini, B. liaoningense, B. yuanmingense, B. subterraneum, B. brasilense and B. retamae. Phylogenetic analysis of the nodC symbiotic gene clustered 13 strains in three different symbiovars (sv. vignae, sv. genistearum and sv. retamae), while seven others might compose new symbiovars. The genetic profiles of the strains evaluated by BOX-PCR revealed high intra- and interspecific diversity. The results point out the high level of diversity still to be explored within the Bradyrhizobium genus, and further studies might confirm new species and symbiovars.     

Mate Recognition in the South African Citrus Thrips Scirtothrips aurantii (Faure) and Cross-Mating Tests with Populations from Australia and South Africa

South African citrus thrips (Scirtothrips aurantii) is a pest of citrus, mango and other horticultural species in its native range, which encompasses a large part of Africa. Its adventitious establishment in Australia in 2002 was a major cause for concern. The thrips, 11 years after its incursion into Australia, has remained on plants of a single host plant genus Bryophyllum (Crassulaceae). Characterization of the Specific-Mate Recognition System of the Bryophyllum population of thrips present in Australia and behavioral bioassay experiments revealed that compounds found in the insects’ body extracts play a crucial role in mate recognition of S. aurantii. Reciprocal cross-mating experiments between the Australian Bryophyllum insects and South African S. aurantii from horticultural host plants showed that mating frequencies were significantly lower in test crosses (Bryophyllum x horticultural) than in controls (Bryophyllum x Bryophyllum or horticultural x horticultural), which indicates there are at least two distinct species within S. aurantii and suggests further tests of this interpretation. The results suggest that these tiny phytophagous insects localize mates through their association with a particular host plant species (or closely-related group of species). Also, specific tests are suggested for clarifying the species status of the host-associated populations of S. aurantii in Africa.     

Expansion of Genetic Diversity in Randomly Mating Founder Populations of Alternaria brassicicola Infecting Cakile maritima in Australia

Founder populations of fungal plant pathogens are expected to have low levels of genetic diversity coupled with further genetic drift due to, e.g., limited host availability, which should result in additional population bottlenecks. This study used microsatellite markers in the interaction between Cakile maritima and the fungal pathogen Alternaria brassicicola to explore genetic expectations associated with such situations. The host, C. maritima, was introduced into Australia approximately 100 years ago, but it is unknown whether the pathogen was already present in Australia, as it has a wide occurrence, or whether it was introduced to Australia on brassicaceous hosts. Eleven A. brassicicola populations were studied, and all showed moderate levels of gene and genotypic diversity. Chi-square tests of the frequencies of mating type alleles, a large number of genotypes, and linkage equilibrium among microsatellite loci all suggest A. brassicicola reproduces sexually. Significant genetic differentiation was found among populations, but there was no evidence for isolation by distance effects. Bayesian analyses identified eight clusters where the inferred clusters did not represent geographical populations but instead consisted of individuals admixed from all populations. Further analysis indicated that fungal populations were more likely to have experienced a recent population expansion than a population bottleneck. It is suggested that A. brassicicola has been introduced into Australia multiple times, potentially increasing the diversity and size of any A. brassicola populations already present there. Combined with its ability to reproduce sexually, such processes appear to have increased the evolutionary potential of the pathogen through recent population expansions.Analyses of population genetic diversity provide useful information on the epidemiology and evolutionary history of infectious diseases (6). The effective population sizes, transmission abilities, and reproduction of infectious agents vary, all of which can be used to infer their population histories (25). Such inferences can in turn be used to determine whether the pathogen has been recently introduced (i.e., a founder population), as well as the number of introductions that have occurred. Introductions of plant pathogens into new geographic areas have occurred commonly in the past for a multitude of organisms, reducing their population diversity in the founder populations but sometimes also causing an epidemic in naïve hosts in the areas of introduction. For example, Phytophthora infestans, a pathogen of the potato, was introduced from Mexico into the United States, and then only one genotype was introduced into Europe (24), causing the death of more than 1.5 million people due to starvation (12). Cryphonectria parasitica, which causes chestnut blight, was introduced from Asia into North America (2), where it underwent a host shift from Asian chestnuts to North American chestnuts, threatening the existence of North American chestnut trees.For both plant and animal pathogens, introduction into new areas can be a major driver of emerging diseases, i.e., host shifts (3, 44, 66). In the case of plant pathogens, range expansion can occur after the introduction of an invasive host species (7, 41), resulting in a pathogen with evolutionary potential higher than that represented in the original founder population. Population genetic expansions for plant pathogens could result in the pathogen being able to infect a larger number of plant genotypes or even new host species following a host shift (38).In agricultural systems, many fungal plant pathogens have genetic structures consistent with having experienced a population bottleneck due to founder events (24, 33, 41, 49); others, such as Rhynchosporium secalis on barley (32) and Mycosphaerella graminicola on wheat (5, 32), eventually underwent a population expansion, possibly due to an increase in host availability. One consequence of this is the accumulation of additional mutations, thereby creating novel genotypes and increasing the effective population size of the pathogen. However, even in well-studied agricultural-plant-pathogen systems, tests for population expansions have rarely been conducted. In most cases where there is evidence for the genetic expansion of pathogen populations, it was a result of additional migrants coming into the founder population (e.g., P. infestans [23]) or the occurrence of sexual reproduction, as in European populations of C. parasitica (41). An increase in genotypic diversity has also been demonstrated in a founder population of the Dutch elm disease pathogen Ophiostomo novo-ulmi in Portugal, which was able to reproduce sexually following the introduction of the other mating type (13). The evolution of founder pathogen populations is likely to be influenced by a range of ecological and life history parameters, including host plant population density, host longevity, and host and pathogen mating systems and dispersal abilities (6, 45).In theory, bottlenecks reduce gene diversity and the number of alleles in populations. Empirically, it has been shown that founder events frequently reduce allelic diversity and, more rarely, gene diversity (15, 21, 28). Thus, we can detect whether a population has experienced a recent population expansion after a demographic bottleneck by using genetic information to determine whether the population has a deficit in gene diversity relative to the number of alleles (16, 35), or whether it experienced a recent population bottleneck, in which case an excess in gene diversity relative to the number of alleles under a mutation drift model will be present (31).In this study, we investigated the introduced Cakile maritima-Alternaria brassicicola host-pathogen interaction in Australia. A. brassicicola is a heterothallic haploid fungus that causes black spots on the leaves, stems, and fruits of a wide range of brassicaceous hosts. The pathogen has a wide distribution and is commonly found on both agricultural brassicaceous crops and wild species (53, 61, 65). The pathogen naturally occurs on C. maritima, a succulent foredune annual native to the Mediterranean and Western Europe that was introduced into Australia more than 100 years ago (50). C. maritima is a self-incompatible, obligate outcrossing species with low levels of self-fertilization (62) and produces seeds that can survive for long periods immersed in seawater. Therefore, seed dispersal between populations is possible, both in terms of seed survival and ocean current patterns (22), and it is estimated that in Australia, C. maritima has spread along the coast at rates of 50 to 100 km per year (51). The conidia (asexual spores) of A. brassicicola are airborne and are dispersed through wind and rain splash, as well as vertically via infected seeds transported by ocean currents. In such interactions, where there is significant potential for seed-borne pathogen transmission, it is expected that the pathogen population should track the dispersal of the host and, to some extent, display a population structure that reflects the host''s geography.Whether A. brassicicola is able to reproduce sexually is still unknown, although it is presumed to be an asexual fungus, as it has no known sexual cycle. However, studies using amplified fragment length polymorphisms (AFLPs) have shown high levels of gene and genotypic diversity in Australian populations of A. brassicicola infecting C. maritima (9, 10). Although the populations were in significant gametic disequilibrium, the authors deemed the levels of gametic disequilibrium to be low and predicted that a sexual stage should be present based on the high genotypic diversity observed (9, 10). Asexual organisms are generally characterized by an overrepresentation of particular genotypes, and therefore, genotype diversity is generally low, whereas recombination of alleles creates numerous new genotypes in sexually reproducing populations (37, 40, 63). It is worth noting, in the context of the preceding discussion regarding pathogen dispersal, that sexual reproduction in A. brassicicola would result in the formation of sexual spores (ascospores), which are smaller than asexual spores, potentially leading to longer-distance wind dispersal.The aims of this study were to further investigate the population genetic structure of A. brassicicola and to determine whether populations are randomly mating. The mating type frequencies of A. brassicicola in either natural or agricultural host populations have never been determined. Random mating was investigated by (i) analyzing levels of genetic diversity and linkage disequilibrium in Alternaria populations characterized with microsatellite markers and (ii) quantifying the extent of frequency-dependent selection of pathogen mating type alleles in these populations. Sexual reproduction can occur in heterothallic fungi only when isolates of opposite mating types are in close physical proximity to each other; therefore, the finding that both mating types occur in a population at equal frequencies (i.e., through frequency-dependent selection of the mating types) would imply that random mating is occurring. Furthermore, we investigated the evolution and genetic structures of fungal populations. Specifically, we investigated whether (i) local populations belong to one panmictic population representing one population; (ii) populations have undergone a recent (in the last 100 years) population bottleneck, typical of founder populations; or (iii) whether populations have expanded genetically, as well as geographically, resulting in pathogen populations with an evolutionary potential higher than that in the original population (38).     

Mechanisms for enhancing nutrient uptake in plants,with particular reference to mediterranean South Africa and Western Australia

The major constraints to nutrient uptake by vascular plants in mediterranean South Africa and Western Australia are: very infertile soils, relatively low temperatures when water availability is high, and hot, dry summers. These constraints are partly overcome through increased efficiency of uptake, tapping novel sources of nutrients, and prolonging water uptake. Absorptive area per unit “cost” may be enlarged directly through increased fineness of the root system and proliferation of long root hairs. This reaches its greatest development in the root clusters of the Proteaceae (proteoid roots), Restionaceae (“capillaroid” roots) and Cyperaceae (dauciform roots). Absorptive area is increased indirectly through fungal hyphae which extend from hairless rootlets into the soil. Two major groups can be recognised: general (VA mycorrhizas) and host-specific (ericoid, orchid and sheathing mycorrhizas). Mycorrhizas are the most widespread specialised modes of nutrition and are probably universal in such major taxa here asPodocarpus, Acacia, Fabaceae, Poaceae, Asteraceae, Rutaceae, terrestrial orchids, Ericales and Myrtaceae. General mycorrhizas are the least drought-adapted of mechanisms for maximising absorptive area. All have been implicated in enhancing P uptake through increasing access to inorganic P, solubilisation and shortening the diffusion path. However, selective uptake of other nutrients, especially N, by host-specific mycorrhizas may be equally important. Included under novel sources of nutrients are free N₂ (utilised by N₂-fixing nodules), small-animal prey (carnivorous leaves) and persistent leaf bases (aerial roots ofKingia australis). Both legume and non-legume N₂-fixing species are well-represented in these two regions, with stands of individual species in southwestern Australia estimated to contribute 2–19 kg N/ha/yr to the ecosystem. Free nitrogen fixation requires additional nutrients, especially Mo and Co, but is enhanced following fires and by supplementary uptake mechanisms, especially VA mycorrhizas. Southwestern Australia is particularly rich in carnivorous species. Nitrogen, P, K and S are important nutrients absorbed, with digestion aided by enzymes provided by bacteria and the glands. Parasitic plants both tap novel sources of nutrients and capitalise on any efficient water and nutrient uptake mechanisms of the hosts. Root parasites are better represented than stem parasites in mediterranean South Africa and Western Australia. Phosphorus and K in particular are absorbed preferentially by the haustoria, but much remains to be known about their modes of operation. Maximum activity of all uptake mechanisms, except those attached to some deep-rooted plants, is restricted to winter-spring. Most new seasons’s rootlets and specialised roots are confined to the uppermost 15 cm of soil, especially in or near the decomposing litter zone. Nutrient uptake is further enhanced by the tendency for the rootlets to cluster, trapping water by capillary action and prolonging nutrient release. As an early product of decomposition, N tends to be available as NH₄ (rather than NO₃) and it is absorbed preferentially by almost all specialised modes of nutrition. Microorganisms are required in the formation and/or functioning of all these structures, except haustoria. Uptake mechanisms which are optional to the plant reach their peak contribution to the root system at soil nutrient levels well below those required for greatest plant growth, when they may be absent altogether. It is only over the narrow range of nutrient availability, where shoot content of a nutrient is greater in the presence of the mechanism than in its absence (other factors remaining constant), that specialised modes can be termed nutrient-uptake “strategies.” For all specialised modes of nutrition, the component genera are better represented in these two regions than in the surrounding more fertile, arid to subtropical regions of much greater area. Endemism of species with each mode exceeds that for the two floras overall (75%). This is taken as preliminary evidence that specialised modes of nutrition are best represented in nutrient-poor soils. While they serve to limit nutrient loss from the ecosystem, their proliferation is therefore not necessarily a response to increasing “leaks” in the system. A hierarchical scheme of the functional/structural relationships between the various mechanisms is presented, starting with the rootless, VA-mycorrhizal plant as the most primitive condition. Taxa with many of the specialised modes of nutrition at present in southwestern South Africa and Western Australia have been evident in the pollen record since the early Tertiary Period. The absence of ectomycorrhizal forests in mediterranean South Africa, in marked contrast to Western Australia, can be traced to differences in their paleohistory. In both regions, the combination of fluctuating, but essentially diminishing, nutrient and water availability that began with the first mediterranean climate < 5 million years ago resulted in decimation of the less-tolerant rainforest ancestors on the one hand, and remarkable rates of speciation of the pre-adapted sclerophyll nucleus on the other.     

Fire-stimulated flowering among resprouters and geophytes in Australia and South Africa

Data on 386 species with fire-stimulated flowering (fsf) in Australasia and South Africa/Madagascar were collated to show that they occur under a wide range of fire regimes, with 71% confined to the mediterranean-climate regions. About 40% only flower up to 2 years after fire (obligate), while the rest continue at a low rate until the next fire (facultative). Peak flowering occurs 5–18 months after fire in the mediterranean regions but at 1–7 months in savannas. Fsf is recorded in 34 families, headed by terrestrial orchids (45% of species), spread throughout the seed-plant phylogeny from cycads to daisies. Tuberous geophytes (essentially orchids) dominate (51%), but other resprouting growth forms include lignotuberous shrubs and forbs, rhizomatous and bunch grasses, leaf succulents, grasstrees, epicormic trees, and hemiparasites. Most have wind-dispersed diaspores (72%), store their diaspores in the soil (93%), and seeds that do not germinate until the next fire (72%). Fsf in association with resprouting takes advantage of optimal resources and minimal competition for growth and reproduction, conditions that favor wind dispersal and maximize the interval for seed accumulation before the next fire and build-up of fire-tolerant organs. Reduced herbivory has little role in accounting for its benefits. The proximal causes of fsf center around cueing factors (direct effects such as ethylene), resource factors (direct and indirect effects, e.g., extra nutrients), and predisposing factors (circumstantial effects, e.g., fire interval). The evolutionary history of fsf has been explored recently in orchids, proteas, blood roots, droseras, and mistletoes and shown to stretch back over a period of at least 50 million years, indicating that flowering in many groups has a long association with fire as an agent of natural selection.     

Bird pollination in South Western Australia: A checklist

At least 560 species of flowering plants distributed in 64 genera and 16 families native to Southern Western Australia are pollinated by birds. This represents 15% of the total flora, and indicates adaptation to bird pollination is a major evolutionary force in this region.     

Comparing ecological restoration in South Africa and Western Australia: the benefits of a ‘travelling workshop’

Researchers and practitioners of ecological restoration from South Africa and Western Australia recently undertook a travelling workshop across several South African biomes to discuss and evaluate the various approaches currently being utilised in the field. Key findings from the workshop highlighted the contrasting nature of ecological restoration undertaken in these two ecologically similar, but socially and politically different regions.     

Diversity among Field Populations of Bradyrhizobium japonicum in Poland

Genetic structure in field populations of Bradyrhizobium japonicum isolated in Poland was determined by using several complementary techniques. Of the 10 field sites examined, only 4 contained populations of indigenous B. japonicum strains. The Polish bradyrhizobia were divided into at least two major groups on the basis of protein profiles on polyacrylamide gels, serological reaction with polyclonal antisera, repetitive extragenic palindromic PCR fingerprints of genomic DNA, and Southern hybridization analyses with nif and nod gene probes. Serological analyses indicated that 87.5% of the Polish B. japonicum isolates tested were in serogroups 123 and 129, while seven (12.5%) of the isolates tested belonged to their own unique serogroup. These seven strains also could be grouped together on the basis of repetitive extragenic palindromic PCR fingerprints, protein profiles, and Southern hybridization analyses. Cluster analyses indicated that the seven serologically undefined isolates were genetically dissimilar from the majority of the Polish B. japonicum strains. Moreover, immuno-cross-adsorption studies indicated that although the Polish B. japonicum strains reacted with polyclonal antisera prepared against strain USDA123, the majority failed to react with serogroup 123- and 129-specific antisera, suggesting that Polish bradyrhizobia comprise a unique group of root nodule bacteria which have only a few antigens in common with strains USDA123 and USDA129. Nodulation studies indicated that members of the serologically distinct group were very competitive for nodulation of Glycine max cv. Nawiko. None of the Polish serogroup 123 or 129 isolates were restricted for nodulation by USDA123- and USDA129-restricting soybean plant introduction genotypes. Taken together, our results indicate that while genetically diverse B. japonicum strains were isolated from some Polish soils, the majority of field sites contained no soybean-nodulating bacteria. In addition, despite the lack of long-term soybean production in Poland, field populations of unique B. japonicum strains are present in some Polish soils and these strains are very competitive for nodulation of currently used Polish soybean varieties.     

Molecular characterisation of Cryptosporidium outbreaks in Western and South Australia

Molecular typing at the 18S rRNA and Gp60 loci was conducted on Cryptosporidium-positive stool samples from cases collected during 2007 Western Australian and South Australian outbreaks of cryptosporidiosis. Analysis of 48 Western Australian samples identified that all isolates were C. hominis and were from five different Gp60C. hominis subtype families. The IbA10G2 subtype was most common across all age groups (37/48). In South Australia, analysis of 24 outbreak samples, identified 21 C. hominis isolates, two C. parvum isolates and one sample with both C. hominis and C. parvum. All C. hominis isolates were identified as the IbA10G2 subtype.     

Dynamics of Introduced Populations of Phragmidium violaceum and Implications for Biological Control of European Blackberry in Australia

Phragmidium violaceum causes leaf rust on the European blackberry (Rubus fruticosus L. aggregate). Multiple strains of this pathogen have been introduced into southern Australia for the biological control of at least 15 taxa of European blackberry, a nonindigenous, invasive plant. In climates conducive to leaf rust, the intensity of disease varies within and among infestations of the genetically variable host. Genetic markers developed from the selective amplification of microsatellite polymorphic loci were used to assess the population genetic structure and reproductive biology of P. violaceum within and among four geographically isolated and diseased infestations of the European blackberry in Victoria, Australia. Despite the potential for long-distance aerial dispersal of urediniospores, there was significant genetic differentiation among all populations, which was not associated with geographic separation. An assessment of multilocus linkage disequilibrium revealed temporal and geographic variation in the occurrence of random mating among the four populations. The presence of sexual spore states and the results of genetic analyses indicated that recombination, and potentially random migration and genetic drift, played an important role in maintaining genotypic variation within populations. Recombination and genetic differentiation in P. violaceum, as well as the potential for metapopulation structure, suggest the need to release additional, genetically diverse strains of the biocontrol agent at numerous sites across the distribution of the Australian blackberry infestation for maximum establishment and persistence.     

Host Association of Cryptosporidium parvum Populations Infecting Domestic Ruminants in Spain

A stock of 148 Cryptosporidium parvum DNA extracts from lambs and goat kids selected from a previous study examining the occurrence of Cryptosporidium species and GP60 subtypes in diarrheic lambs and goat kids in northeastern Spain was further characterized by a multilocus fragment typing approach with six mini- and microsatellite loci. Various degrees of polymorphism were seen at all but the MS5 locus, although all markers exhibited two major alleles accounting for more than 75% of isolates. A total of 56 multilocus subtypes (MLTs) from lambs (48 MLTs) and goat kids (11 MLTs) were identified. Individual isolates with mixed MLTs were detected on more than 25% of the farms, but most MLTs (33) were distinctive for individual farms, revealing the endemicity of cryptosporidial infections on sheep and goat farms. Comparison with a previous study in calves in northern Spain using the same six-locus subtyping scheme showed the presence of host-associated alleles, differences in the identity of major alleles, and very little overlap in MLTs between C. parvum isolates from lambs and those from calves (1 MLT) or isolates from lambs and those from goat kids (3 MLTs). The Hunter-Gaston index of the multilocus technique was 0.976 (95% confidence interval [CI], 0.970 to 0.982), which supports its high discriminatory power for strain typing and epidemiological tracking. Population analyses revealed the presence of two host-associated subpopulations showing epidemic clonality among the C. parvum isolates infecting calves and lambs/goat kids, respectively, although evidence of genetic flow between the two subpopulations was also detected.     

Fractionation and Estimation of Particle-Attached and Unattached Bradyrhizobium japonicum Strains in Soils

Rhizobial cells attached or unattached to soil particles were estimated. Nonsterile soils into which antibiotic-resistant mutants of Bradyrhizobium japonicum had been introduced were fractionated by a centrifugation technique into two fractions: A, which contained mainly rhizobial cells attached to soil particles, and F, which contained mainly rhizobial cells unattached to them. Rhizobial counts decreased in both fractions during incubation of the soil at 30°C, with a concomitant decrease in the proportion of the count of fraction F to that of fraction A. Sonication of fraction A of the soil incubated for more than 3 weeks caused an increase in the rhizobial count. The ratio of the count of fraction A estimated by the plant infection method to that estimated by the dilution plate method increased after 5 days of soil incubation. More than 90% of the indigenous rhizobia in an agricultural field existed in fraction A. These results suggest that the majority of rhizobial cells are attached to soil particles.     

Prenatal Exposure to Cadmium,Placental Permeability and Birth Outcomes in Coastal Populations of South Africa

Background

The impact of prenatal exposure to cadmium (Cd) on birth outcomes is an area of concern. This study aimed to assess an impact of prenatal Cd exposure on birth outcomes in distinct coastal populations of South Africa.

Methods

Cadmium was measured in maternal blood (CdB) (n = 641), cord blood and in maternal urine (n = 317). This investigation assessed the associations between CdB (non-transformed) and birth outcomes across the 25^th, 50th, and 75th percentile for birth weight, birth length and head circumference, to test for a linear trend. Associations between natural log-transformed maternal CdB, size at birth and other factors were further evaluated using linear mixed-effects modelling with random intercepts.

Results

The average gestational age in the total sample was 38 weeks; 47% of neonates were female, average birth weight was 3065 g and 11% were of low birth weight (< 2500 g). The geometric mean (GM) of the maternal CdB level was 0.25 μg/L (n = 641; 95% CI, 0.23–0.27). The cord blood Cd level was 0.27 μg/L (n = 317; 95% CI, 0.26–0.29) and urine (creatinine-corrected) Cd level was 0.27 μg/L (n = 318; 95% CI, 0.24–0.29). The CdB cord:maternal ratio in the sub-cohort was 1, suggesting that the placenta offers no protective mechanism to the foetus. An inverse association was found between CdB and the lower birth weight percentile in female neonates only (β = - 0.13, p = 0.047). Mothers who reported eating vine vegetables daily had lower levels of CdB (β = - 0.55, p = 0.025). Maternal smoking was associated with an elevation in natural log-transformed CdB levels in both male and female cohorts.

Discussion

Significant inverse associations between prenatal Cd exposure and birth anthropometry were found in female neonates but not in male neonates, suggesting potential sex differences in the toxico-kinetics and toxico-dynamics of Cd.     

Bradyrhizobium xenonodulans sp. nov. isolated from nodules of Australian Acacia species invasive to South Africa

A genealogical concordance approach was used to delineate strains isolated from Acacia dealbata and Acacia mearnsii root nodules in South Africa. These isolates form part of Bradyrhizobium based on 16S rRNA sequence similarity. Phylogenetic analysis of six housekeeping genes (atpD, dnaK, glnII, gyrB, recA and rpoB) confirmed that these isolates represent a novel species, while pairwise average nucleotide identity (ANIb) calculations with the closest type strains (B. cosmicum 58S1^T, B. betae PL7HG1^T, B. ganzhouense CCBAU 51670 ^T, B. cytisi CTAW11^T and B. rifense CTAW71^T) resulted in values well below 95–96%. We further performed phenotypic tests which revealed that there are high levels of intraspecies variation, while an additional analysis of the nodA and nifD loci indicated that the symbiotic loci of the strains are closely related to those of Bradyrhizobium isolates with an Australian origin. Strain 14AB^T (=LMG 31415 ^T = SARCC-753 ^T) is designated as the type strain of the novel species for which we propose the name Bradyrhizobium xenonodulans sp. nov.     

Differentiating Indigenous Soybean Bradyrhizobium and Rhizobium spp. of Indian Soils

Soybean is extensively cultivated worldwide and is the largest source of biologically fixed nitrogen among legumes. It is nodulated by both slow and fast growing rhizobia. Indigenous soybean rhizobia in Vertisols of central India were assessed for utilization of 35 carbon sources and intrinsic resistance to 19 antibiotics. There was greater utilization of trehalose and raffinose by fast growers (87 and 73 % by fast vs. 35 and 30 % by slow growers); but slow growers had higher ability to utilize glucosamine (75 % by slow vs. 33 % by fast growers). A larger proportion of slow growers were resistant to vancomycin, polymyxin-B and rifampicin (70, 65 and 55 %) compared to fast growers (13, 7 and 7 % each). Among the two 16S rRNA sequence types in the slow growers, those belonging to Bradyrhizobium spp. utilized glucosamine while those belonging to Rhizobium radiobacter did not. All the fast growers had 16S rRNA homology to R. radiobacter and majority could not utilize glucosamine. It is suggested that during initial isolations and screening of rhizobia in strain selection programmes, using carbon sources like glucosamine and antibiotics like vancomycin, polymyxin-B and rifampicin in the media may provide a simple way of distinguishing Bradyrhizobium strains from R. radiobacter among the slow growers.