High-Speed Cell Sorting of Infectious Trophic and Cystic Forms of Pneumocystis carinii

ABSTRACT. The separation of Pneumocystis carinii life-cycle stages while preserving infectivity is a hitherto unresolved challenge. We describe an original, reproducible, and efficient method for separating trophic from cystic forms of P. carinii using a high-speed cell sorter. The large amounts of highly purified (99.6±0.3%) infectious trophic and cystic forms can now be used to elucidate the poorly understood P. carinii life cycle.     

Potentiality of Different Seed Priming Agents to Mitigate Cold Stress of Winter Rice Seedling

Seed priming has proved to be an effective pre-germination seed invigoration technique for different crops to improve seed and seedling performance under different abiotic stresses. In Bangladesh, winter rice is very often exposed to cold waves just after sowing in the nursery bed resulting in poor seed germination and seedling emergence, yellowish and thin seedlings production, and a very low survival rate. Seed priming may mitigate the cold stress during seed germination and seedling emergence and helps in the quality seedling production of winter rice. To evaluate the efficacy of different seed priming techniques in increasing seedling emergence, growth, vigor and survivability of winter rice cultivars under cold stress, a pot experiment was conducted at the Department of Agronomy, Bangladesh Agricultural University during December 2018 to January 2019. The experiment comprised two factors, (A) Winter rice variety namely, i) BRRI dhan29 and ii) BRRI dhan36; (B) Seed priming agent namely i) Control (no priming), ii) 20000 ppm NaCl, iii) 30000 ppm NaCl, iv) 20000 ppm KCl, v) 30000 ppm KCl, vi) 20000 ppm CaCl₂, vii) 30000 ppm CaCl₂, viii) 50 ppm CuSO₄, ix) 75 ppm CuSO₄, x) 10000 ppm ZnSO₄, xi) 15000 ppm ZnSO₄, xii) 2 ppm Na₂MoO₄, xiii) 3 ppm Na₂MoO₄, xiv) 100 ppm PEG (Polyethylene glycol 4000) and xv) 150 ppm PEG. Seeds were sown on two different dates viz., 1st December and 1st January so that seedlings are exposed to cold stress at different stages. The experiment was laid out in a completely randomized design (CRD) with three replications. Results indicated that (in most of the cases) seed priming has a positive impact on seedling emergence rate (%), root length, shoot length, root shoot ratio, root dry weight, shoot dry weight, seedling dry weight and survival rate (%). Among the priming agents, KCl and CaCl₂ performed best; while priming with NaCl and PEG showed no advantages over no priming for both the sowing dates. In general, BRRI dhan36 performed better than BRRI dhan29 in terms of seedling growth because of its higher tolerance to cold stress. But, both the varieties performed similarly in terms of emergence rate and survival rate. Thus, priming is an effective tool to increase seed germination, better seedling growth, and higher seedling survivability of winter rice under cold stress, and KCl (20000 ppm) or CaCl₂ (20000 ppm) can be considered as a viable priming agent.     

Comparative analysis of storage conditions and homogenization methods for tick and flea species for identification by MALDI‐TOF MS

Ticks and fleas are vectors for numerous human and animal pathogens. Controlling them, which is important in combating such diseases, requires accurate identification, to distinguish between vector and non‐vector species. Recently, matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF‐MS) was applied to the rapid identification of arthropods. The growth of this promising tool, however, requires guidelines to be established. To this end, standardization protocols were applied to species of Rhipicephalus sanguineus (Ixodida: Ixodidae) Latreille and Ctenocephalides felis felis (Siphonaptera: Pulicidae) Bouché, including the automation of sample homogenization using two homogenizer devices, and varied sample preservation modes for a period of 1–6 months. The MS spectra were then compared with those obtained from manual pestle grinding, the standard homogenization method. Both automated methods generated intense, reproducible MS spectra from fresh specimens. Frozen storage methods appeared to represent the best preservation mode, for up to 6 months, while storage in ethanol is also possible, with some caveats for tick specimens. Carnoy's buffer, however, was shown to be less compatible with MS analysis for the purpose of identifying ticks or fleas. These standard protocols for MALDI‐TOF MS arthropod identification should be complemented by additional MS spectrum quality controls, to generalize their use in monitoring arthropods of medical interest.     

Differences in Activity Budgets and Diet Between Semiprovisioned and Wild‐Feeding Groups of the Endangered Barbary Macaque (Macaca sylvanus) in the Central High Atlas Mountains,Morocco

The Barbary macaque, Macaca sylvanus is a very adaptable primate species occupying a wide range of habitats in Morocco and Algeria. Several groups of this endangered macaque can be found in tourist sites, where they are affected by the presence of visitors providing food to them. We compare the activity budgets and the diet of semiprovisioned and wild‐feeding groups of Barbary macaques in the central High Atlas Mountains of Morocco from February to August 2008. We used instantaneous scan sampling at 15‐min intervals. The behaviors included in the activity budget were feeding, moving, foraging, resting, and aggressive display. Food items were grouped into seven categories. We found no differences between the two groups in the daily percentages of records attributed to feeding. The semiprovisioned group spent significantly more time engaged in resting and aggressive behavior, and foraged and moved significantly less than the wild‐feeding group. There was no significant difference between the two groups in time spent eating leaves, fruits, or roots and bark. The semiprovisioned group, however, spent significantly less time per day feeding on herbs, seeds, and acorns than the wild‐feeding group. Human food accounted for 26% of the daily feeding records for the semiprovisioned group and 1% for the wild‐feeding group. Our findings agree with previous studies and indicate that in the tourist site, where food is highly clumped, macaques decreased foraging time yet showed higher levels of contest competition. Our results support the common claim that the diet of the Barbary macaque is highly flexible, differing among its varied habitats. Conservation efforts for the Barbary macaques should take into account the changes in behavior that human‐modified environments may cause.     

Isolation and characterization of microsatellite markers in the East African tree,Acacia brevispica (Fabaceae: Mimosoideae)

We have developed primer pairs for 24 microsatellite loci isolated from Acacia brevispica, which is widespread in woodland savannas of East Africa. The loci were screened for levels of variation using 16–52 individuals from Mpala Research Centre, Kenya. Number of alleles per locus ranged from two to 17, and polymorphic information content ranged from 0.248 to 0.864. Several loci showed the presence null alleles, but many loci will be useful in ongoing studies of genetic structure, gene flow, breeding systems, and natural selection.     

Molecular cytogenetics studies in Reichardia tingetana: Physical mapping of heterochromatin,telomere repeats,and 5S and 45S rDNA by 4′, 6‐diamidino‐2‐phenylindole and fluorescence in situ hybridization

Abstract Molecular cytogenetics studies of A‐T‐rich regions, telomeres, and 5S and 45S rDNA sites on the chromosomes of Reichardia tingetana Roth (2n= 16; diploid) were done using 4′, 6‐diamidino‐2‐phenylindole (DAPI) and fluorescence in situ hybridization (FISH). The species were collected from three geographically isolated populations at Borg El Arab (salt marsh habitat), and Rashed and Shosha (sandy clay habitats) in Egypt. The three populations showed the chromosome number of all plants are diploid except for two tetraploid samples from Shosha. Plants from both Rashed and Shosha showed similarity in the distribution of six DAPI bands on six chromosomes, whereas those of Borg El Arab showed a distribution of 16 bands on 14 chromosomes. The FISH signals of the telomeres, and 5S and 45S rDNA, were at the telomeres of all chromosomes, two interstitial, and four terminal, respectively. The combination of DAPI and FISH showed colocalization of the DAPI bands with two 5S and two 45S rDNA loci. The increased number of DAPI bands in the cytotypes from the salt marsh habitat could indicate natural genetic adaptation through increasing the heterochromatin of A‐T‐rich regions.     

<Emphasis Type="Italic">Dunaliella salina</Emphasis> exopolysaccharides: a promising biostimulant for salt stress tolerance in tomato (<Emphasis Type="Italic">Solanum lycopersicum</Emphasis>)

Microalgal exopolysaccharides represent a potential sustainable alternative for the enhancement and protection of agricultural crops including management of both biotic and abiotic stress. In the present study, we investigated the potential of Dunaliella salina exopolysaccharides (PS) to attenuate the effect of salt stress on growth of Solanum lycopersicum, which was grown under different salinity levels (3 and 6 g L^?1 NaCl). The effects of PS treatment on plant growth, osmoprotectant molecules, protein content, and antioxidant enzymes activities of tomato plants under salt stress were analyzed. A metabolomics study showed that the exopolysaccharides released by D. salina contained sulfated moiety along with carbohydrates and uronic acids. The application of sulfated exopolysaccharides on tomato plants alleviated the salt stress and mitigated the decrease in length and dry weight of the plant’s shoot and root systems, as well as that of potassium (K⁺), and K⁺/Na⁺ ratio. Furthermore, the increase in proline, phenolic compounds, Na⁺, and antioxidant enzymes (CAT, POD, SOD) activities caused by salt stress were attenuated after the exopolysaccharide treatment. GC-MS metabolomics analysis showed that PS treatment allowed the activation and/or inhibition of various metabolic pathways involved in the plant’s tolerance to stress such as jasmonic acid-dependent pathways. This study shows the potential of microalgal exopolysaccharides for enhancing tomato tolerance to salt stress and highlights the possibility of their use as plant growth biostimulants under harsh environmental conditions.