Bemisia tabaci is one of the most threatening pests in agriculture, causing significant losses to many important crops on a global scale. The dramatic increase and availability of sequence data for B. tabaci species complex and its bacterial endosymbionts is critical for developing emerging sustainable pest management strategies which are based on pinpointing the global diversity of this important pest and its bacterial endosymbionts. To unravel the global genetic diversity of B. tabaci species complex focusing on its associated endosymbionts, along with Israeli whitefly populations collected in this study, we combined available sequences in databases, resulting in a total of 4,253 mitochondrial cytochrome oxidase I (mtCOI) sequences from 82 countries and 1,226 16S/23S rRNA endosymbiont sequences from 32 countries that were analyzed. Using Bayesian phylogenetic analysis, we identified two new B. tabaci groups within the species complex and described the global distribution of endosymbionts within this complex. Our analyses revealed complex divergence of the different endosymbiont sequences within the species complex, with overall one Hamiltonella, two Porteria (P1 and P2), two Arsenophonus (A1 and A2), two Wolbachia (super-groups O and B), four Cardinium (C1-C4) and three Rickettsia (R1-R3) groups were identified. Our comprehensive analysis provides an updated important resource for this globally important pest and its secondary symbionts, which have been a major subject for research in last three decades.

/pdf/global-genetic-diversity-and-geographical-distribution-of-1zun8c7jd2.pdf

Global genetic diversity and geographical distribution of Bemisia tabaci and its bacterial endosymbionts.

The resident microbiota of animals represents an important contribution to the global microbial diversity, but it is poorly known in many animals. This study investigated the bacterial diversity in plant phloem-sap-feeding whiteflies, aphids and psyllids by pyrosequencing bacterial 16S rRNA gene amplicons. After correction for sequencing error, just 3-7 bacterial operational taxonomic units were recovered from each insect sample sequenced to sufficient depth for saturation of rarefaction curves. Most samples were dominated by primary and secondary symbionts, which are localized to insect cells or the body cavity, indicative of a dearth of bacterial colonists of the gut lumen. Diversity indices of the bacterial communities (Shannon's index: 0.40-1.46, Simpson's index: 0.15-0.74) did not differ significantly between laboratory and field samples of the phloem-feeding insects, but were significantly lower than in drosophilid flies quantified by the same methods. Both the low bacterial content of the phloem sap diet and biological processes in the insect may contribute to the apparently low bacterial diversity in these phloem-feeding insects.

The bacterial communities in plant phloem‐sap‐feeding insects

Wolbachia is the most prevalent symbiont described in arthropods to date. Wolbachia can manipulate host reproduction, provide nutrition to insect hosts and protect insect hosts from pathogenic viruses. So far, 13 supergroups of Wolbachia have been identified. The whitefly Bemisia tabaci is a complex containing more than 28 morphologically indistinguishable cryptic species. Some cryptic species of this complex are invasive. In this study, we report a comprehensive survey of Wolbachia in B. tabaci and its relative B. afer from 1658 insects representing 54 populations across 13 provinces of China and one state of Australia. Based on the results of PCR or sequencing of the 16S rRNA gene, the overall rates of Wolbachia infection were 79.6% and 0.96% in the indigenous and invasive Bemisia whiteflies, respectively. We detected a new Wolbachia supergroup by sequencing five molecular marker genes including 16S rRNA, groEL, gltA, hcpA, and fbpA genes. Data showed that many protein-coding genes have limitations in detecting and classifying newly identified Wolbachia supergroups and thus raise a challenge to the known Wolbachia MLST standard analysis system. Besides, the other Wolbachia strains detected from whiteflies were clustered into supergroup B. Phylogenetic trees of whitefly mitochondrial cytochrome oxidase subunit I and Wolbachia multiple sequencing typing genes were not congruent. In addition, Wolbachia was also detected outside the special bacteriocytes in two cryptic species by fluorescence in situ hybridization, indicating the horizontal transmission of Wolbachia. Our results indicate that members of Wolbachia are far from well explored.

https://onlinelibrary.wiley.com/doi/pdf/10.1002/ece3.1126

Diversity and evolution of the Wolbachia endosymbionts of Bemisia (Hemiptera: Aleyrodidae) whiteflies

Vitellogenin (Vg) plays vital role in oocytes and embryo development in insects. Vg is synthesized in the fat body, moves through haemolymph and accumulates in oocytes. Vitellogenin receptors (VgR) present on the surface of oocytes, are responsible for Vg transportation from haemolymph to oocytes. Here, we cloned and characterized these genes from Bemisia tabaci Asia1 (BtA1) species. The cloned BtA1Vg and BtA1VgR genes consisted of 6,330 and 5,430 bp long open reading frames, which encoded 2,109 and 1,809 amino acid (AA) residues long protein. The BtA1Vg protein comprised LPD_N, DUF1943 and VWFD domains, typical R/KXXR/K, DGXR and GL/ICG motifs, and polyserine tracts. BtA1VgR protein contained 12 LDLa, 10 LDLb and 7 EGF domains, and a trans-membrane and cytoplasmic region at C-terminus. Phylogenetic analyses indicated evolutionary association of BtA1Vg and BtA1VgR with the homologous proteins from various insect species. Silencing of BtA1VgR by siRNA did not affect the transcript level of BtA1Vg. However, BtA1Vg protein accumulation in oocytes was directly influenced with the expression level of BtA1VgR. Further, BtA1VgR silencing caused significant mortality and reduced fecundity in adult whiteflies. The results established the role of BtA1VgR in transportation of BtA1Vg in oocytes. Further, these proteins are essential for fecundity, and therefore these can be potential RNAi targets for insect control in crop plants.

/pdf/molecular-characterization-of-vitellogenin-and-vitellogenin-1j5a5frxmj.pdf

Molecular Characterization of Vitellogenin and Vitellogenin Receptor of Bemisia tabaci.

This study was conducted to determine the presence of Acinetobacter and Rickettsia species DNA in lice and Melophagus ovinus (sheep ked) of animals from Oromia Regional State in Ethiopia. From September through November 2011, a total of 207 cattle, 85 sheep, 47 dogs and 16 cats were examined for ectoparasites. Results of morphological identification revealed several species of ectoparasites: Linognathus vituli (L. vituli), Bovicola bovis (B. bovis) and Solenopotes capillatus (S. capillatus) on cattle; B. ovis and Melophagus ovinus (M. ovinus) on sheep; and Heterodoxus spiniger (H. spiniger) on dogs. There was a significantly (p≤0.0001) higher prevalence of L. vituli observed in cattle than both S. capillatus and B. bovis. Molecular identification of lice using an 18S rRNA gene analysis confirms the identified lice species by morphological methods. We detected different Acinetobacter species among lice (11.1%) and keds (86.4%) including A. soli in L. vituli of cattle, A. lowffii in M. ovinus of sheep, A. pittii in H. spiniger of dogs, 1 new Acinetobacter spp. in M. ovinus and 2 new Acinetobacter spp. in H. spiniger of dogs using partial rpoB gene sequence analysis. There was a significantly higher prevalence of Acinetobacter spp. in keds than in lice (p≤0.00001). Higher percentage of Acinetobacter spp. DNA was detected in H. spiniger than in both B. ovis and L. vituli (p≤0.00001). Carbapenemase resistance encoding genes for blaOXA-23, blaOXA-24, blaOXA-58, blaNDM-1 and blaOXA-51 were not found in any lice and keds. These findings suggest that synanthropic animals and their ectoparasites might increase the risk of human exposure to zoonotic pathogens and could be a source for Acinetobacter spp. infections in humans. However, additional epidemiological data are required to determine whether ectoparasites of animals can act as environmental reservoirs and play a role in spreading these bacteria to both animal and human hosts.

/pdf/molecular-detection-of-acinetobacter-species-in-lice-and-3p6nr56o6i.pdf

Molecular detection of Acinetobacter species in lice and keds of domestic animals in Oromia Regional State, Ethiopia.

Diversity and phylogenetic analysis of endosymbiotic bacteria from field caught Bemisia tabaci from different locations of North India based on 16S rDNA library screening

Partially balanced crossover designs (PBCODs) are needed in case a balanced design is not possible due to some reasons. Blaisdell and Raghavarao (1980) and Raghavarao and Blaisdell (1985) introduced the concept of PBCODs and gave certain classes of such designs, along with their efficiency factors. The work on the construction of PBCODs was further considered by Aggarwal and Jha (2006), wherein they gave a number of classes of such designs, based on certain partially balanced incomplete block (PBIB) (2) designs and rectangular association scheme. The present work provides some new classes of connected PBCODs based on 2-, 3-, 4-, and general m-associate class association schemes.

Constructions of partially balanced crossover designs based on two and higher order association schemes

In cross-over designs, sequences of treatments are applied to the units, spread over a specified period of time. The model for the estimation of such designs includes a carryover term, which makes them competent for use in consumer research trials. Construction of useful cross-over designs in the context of consumer research trials forms the body of this work and some new classes of partially balanced cross-over designs with two associate classes [PBCOD(2)] are obtained, along with their efficiencies. Designs constructed are uniform over periods for a large number of units and they estimate treatment contrasts with two sets of precision.

/pdf/partially-balanced-cross-over-designs-for-consumer-trials-3a86v3gxk3.pdf

Partially balanced cross-over designs for consumer trials

In medical research, sequences of treatments may need to be applied to subjects over a period of time. Cross-over designs, in such cases, form a useful class for application. However, there may be some additional factor of variation embedded into the subjects, or the required constraints for application of a balanced design, (which is preferred), may not always be available and the cross-over designs may need to be conducted in parts or sessions. Nested cross-over designs are required under such conditions. Some such situations are considered here and useful nested cross-over designs are constructed. Relevant theory on modeling the observations obtained from such designs is also provided.

Nested cross-over designs

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Diversity and phylogenetic analysis of endosymbiotic bacteria from field caught Bemisia tabaci from different locations of North India based on 16S rDNA library screening

Constructions of partially balanced crossover designs based on two and higher order association schemes

Partially balanced cross-over designs for consumer trials

Nested cross-over designs