Characterization of Vibrio cholerae O1 isolates responsible for cholera outbreaks in Kenya between 1975 and 2017

Kenya is endemic for cholera with different waves of outbreaks having been documented since 1971. In recent years, new variants of Vibrio cholerae O1 have emerged and have replaced most of the traditional El Tor biotype globally. These strains also appear to have increased virulence, and it is important to describe and document their phenotypic and genotypic traits. This study characterized 146 V. cholerae O1 isolates from cholera outbreaks that occurred in Kenya between 1975 and 2017. Our study reports that the 1975–1984 strains had typical classical or El Tor biotype characters. New variants of V. cholerae O1 having traits of both classical and El Tor biotypes were observed from 2007 with all strains isolated between 2015 and 2017 being sensitive to polymyxin B and carrying both classical and El Tor type ctxB. All strains were resistant to Phage IV and harbored rstR, rtxC, hlyA, rtxA and tcpA genes specific for El Tor biotype indicating that the strains had an El Tor backbone. Pulsed field gel electrophoresis (PFGE) genotyping differentiated the isolates into 14 pulsotypes. The clustering also corresponded with the year of isolation signifying that the cholera outbreaks occurred as separate waves of different genetic fingerprints exhibiting different genotypic and phenotypic characteristics. The emergence and prevalence of V. cholerae O1 strains carrying El Tor type and classical type ctxB in Kenya are reported. These strains have replaced the typical El Tor biotype in Kenya and are potentially more virulent and easily transmitted within the population.     

Biological effects of salinity gradient reversals in a southeast African estuarine lake

St Lucia Lake on the north coast of Natal, South Africa, has an area of 325 km² and is the largest estuarine complex in Africa. It consists of a 20 km tidal channel, averagingca. 400 m in width, linking the sea with the non-tidal lake which is H-shaped with a maximum length ofca. 40 km and width ofca. 20 km. Except during flood periods the depth of the lake does not exceed 2 m. The salinity gradient depends on evaporation, the configuration of the mouth and on the input of fresh water from four rivers which discharge into the northern and western areas of the lake. If fresh water input is high, the lake and much of the channel may be fresh. An intermediate stage features a normal salinity gradient while a third stage shows a reversed salinity gradient with salinities in excess of 100‰ in the upper reaches of the system. Changing salinities have marked effects on the biota. Aquatic macrophytes show cycles of appearance and disappearance depending on salinity tolerance and the presence of dormant stages. The resident benthic faunal species go through cycles of range expansion and contraction depending on prevailing salinities and recolonisation by dispersal phases. To date salinities in the southern part of the lake have approached, but not exceeded, lethal levels and this has therefore acted as a reservoir area. Catchment degradation and water abstraction are anticipated to exacerbate future salinity extremes. This has resulted in concern for the long term viability of this Ramsar site which has major southern African populations of hippopotamus and crocodile, provides breeding sites for South African Red Data water bird species and plays an important nursery role for marine fish and penaeid prawns.     

Polyamine derivatives as inhibitors of trypanothione reductase and assessment of their trypanocidal activities

Trypanothione reductase (TR) occurs exclusively in trypanosomes and leishmania, which are the etiological agents of many diseases. TR plays a vital role in the antioxidant defenses of these parasites and inhibitors of TR have potential as antitrypanosomal agents. We describe the syntheses of several spermine and spermidine derivatives and the inhibiting effects of these compounds on T. cruzi TR. All of the inhibiting compounds displayed competitive inhibition of TR-mediated reduction of trypanothione disulfide. The three most effective compounds studied were N⁴,N⁸-bis(3-phenylpropyl)spermine (12), N⁴,N⁸-bis(2-naphthylmethyl)spermine (14), and N¹,N⁸-bis(2-naphthylmethyl)spermidine (21), with K_i values of 3.5, 5.5 and 9.5 μM, respectively. Compounds 12, 14, and 21 were found to be potent trypanocides in vitro with IC₅₀ values ranging from 0.19 to 0.83 μM against four T. brucei ssp. strains. However, these compounds did not prolong the lives of mice infected with trypanosomes. This work indicates that certain polyamine derivatives which target a unique pathway in Trypanosomatidae have potential as antitrypanosomal agents.     

The influence of zooplankton food resources on the morphology of the estuarine clupeid Gilchristella aestuarius (Gilchrist, 1914)

Synopsis The body shape of the estuarine clupeidGilchristella aestuarius from the St. Lucia system is different from that of the same species in other estuaries. The morphology ofG. aestuarius is discussed in relation to long term food availability in the St. Lucia system.     

Effects of Cycloate on Development of Heterodera schachtii and Growth of Three Beta Species

Greenhouse tests were set up to evaluate the effects of the herbicide, cycloate (S-ethyl cydohexylethylthiocarbamate), oil development of Heterodera schachtii and growth of three Beta species. Cycloate added to infested soil enhanced cyst development/gm root on B. vulgaris and larvae/gm of root in B. patellaris and B. procumbens at 4, 16, and 16 μg(a.i.)/gm of soil, respectively. Total numbers of nematodes/individual root system decreased because of poor root growth of seedlings in cycloate-amended soil. Penetration and larval development through stage three did occur in the wild Beta species in any treatment. Thus, resistance of B. patellaris and B. pocumbens to development of H. schachtii was not altered by cycloate. Cycloate also retarded growth (P = 0.05) of the sugarbeet cultivars and B. patellaris at 4 μg(a.i.)/gm and B. procumbens at 16 μg(a.i.)/gm of soil. Higher concentrations of nematodes/gm root in plants growing in cycloate-amended soil may be attributed to factors such as fewer roots available for penetration, possible effects of cycloate on egg hatch, greater attraction of nematodes to roots, and increased susceptibility of roots to larval penetration. Suppression of seedling growth in cycloate-amended soil may be attributed in part to higher nematode density and in part to direct root damage from cycloate.     

Multivariate data analysis of growth medium trends affecting antibody glycosylation

Use of multivariate data analysis for the manufacturing of biologics has been increasing due to more widespread use of data-generating process analytical technologies (PAT) promoted by the US FDA. To generate a large dataset on which to apply these principles, we used an in-house model CHO DG44 cell line cultured in automated micro bioreactors alongside PAT with four commercial growth media focusing on antibody quality through N-glycosylation profiles. Using univariate analyses, we determined that different media resulted in diverse amounts of terminal galactosylation, high mannose glycoforms, and aglycosylation. Due to the amount of in-process data generated by PAT instrumentation, multivariate data analysis was necessary to ascertain which variables best modeled our glycan profile findings. Our principal component analysis revealed components that represent the development of glycoforms into terminally galacotosylated forms (G1F and G2F), and another that encompasses maturation out of high mannose glycoforms. The partial least squares model additionally incorporated metabolic values to link these processes to glycan outcomes, especially involving the consumption of glutamine. Overall, these approaches indicated a tradeoff between cellular productivity and product quality in terms of the glycosylation. This work illustrates the use of multivariate analytical approaches that can be applied to complex bioprocessing problems for identifying potential solutions.     

Biochemical characterization of an E. coli cell division factor FtsE shows ATPase cycles similar to the NBDs of ABC-transporters

The peptidoglycan (PG) layer is an intricate and dynamic component of the bacterial cell wall, which requires a constant balance between its synthesis and hydrolysis. FtsEX complex present on the inner membrane is shown to transduce signals to induce PG hydrolysis. FtsE has sequence similarity with the nucleotide-binding domains (NBDs) of ABC transporters. The NBDs in most of the ABC transporters couple ATP hydrolysis to transport molecules inside or outside the cell. Also, this reaction cycle is driven by the dimerization of NBDs. Though extensive studies have been carried out on the Escherchia coli FtsEX complex, it remains elusive regarding how FtsEX complex helps in signal transduction or transportation of molecules. Also, very little is known about the biochemical properties and ATPase activities of FtsE. Because of its strong interaction with the membrane-bound protein FtsX, FtsE stays insoluble upon overexpression in E. coli, and thus, most studies on E. coli FtsE (FtsE_Ec) in the past have used refolded FtsE. Here in the present paper, for the first time, we report the soluble expression, purification, and biochemical characterization of FtsE from E. coli. The purified soluble FtsE exhibits high thermal stability, exhibits ATPase activity and has more than one ATP-binding site. We have also demonstrated a direct interaction between FtsE and the cytoplasmic loop of FtsX. Together, our findings suggest that during bacterial division, the ATPase cycle of FtsE and its interaction with the FtsX cytoplasmic loop may help to regulate the PG hydrolysis at the mid cell.     

FISH CONSERVATION IN SOUTH AFRICAN ESTUARIES: PRESSURES,PROBLEMS AND PROSPECTS

Summary

A review of the conservation status of fish in the estuarine environment around the South African coastline reveals that some species face serious problems associated either with habitat destruction, and its associated biological, physical and chemical components or exploitation. The 65 species considered fall into three categories; truly estuarine species, which are dependent on estuaries for their entire life; marine species dependent on estuaries during the juvenile phase of their life cycle; and marine species whose juveniles occur mainly in estuaries but are also found at sea. Included in the first two categories are 14 species of fish which are on the South African Red Data list. These comprise one species which is endangered, five which are vulnerable and eight which are rare. All groups are considered in relation to factors in estuaries which are affecting their populations. A conservation strategy is suggested for certain estuarine types or for specific estuaries which could ensure the survival of the entire range of estuarine faunas.     

THE LAKE ST. LUCIA SYSTEM—A RESEARCH ASSESSMENT

SUMMARY

This paper assesses the research undertaken at Lake St. Lucia over the past 25 years based on over 300 documents from that period. Trends related to both time and subject matter are evident, and these are considered in relation to the gaps in our current knowledge concerning the system.

A feature evident throughout the period under consideration is that the major portion of documented material available relates to reports and contributions to workshops (77%) with only 23% from scientific publications. Contributions by these two sources to the subject group being considered for St. Lucia are markedly different with research publications dominating the biological field and reports dominating in physical aspects, catchment characteristics, man's activities, management, dredging and hydrological modelling. However, some 55% of all unpublished data related to reviews or assessments of the state of research on St. Lucia.

The importance of the scientific publications group as an indicator of the state of research into the system is considered in the light of an apparent decline in the number of completed projects being published. It is also considered in the light of the recent establishment of a co-ordinated Lake St. Lucia Research Programme, which may provide the impetus for a more concentrated and directed research effort on the Lake System.