Effect of macrophage secretory products on elaboration of virulence factors by planktonic and biofilm cells of Pseudomonas aeruginosa

Macrophages, which constitute the first line of defense, pour their secretions in the mileu following stimulation with pathogens. These secretory products, referred to as macrophage secretory products (MSPs), can influence ultimate outcome of an infection. In the present investigation, it was observed that different strains of Pseudomonas aeruginosa vary in their ability to stimulate macrophages leading to variability in generation of macrophage secretory products. Cytokine levels, reactive nitrogen intermediates and protein content of macrophage secretory products generated with biofilm cells of P. aeruginosa was found to be more as compared to their planktonic counterparts. The effect of macrophage secretory products produced in response to interaction of macrophages with P. aeruginosa on elaboration of virulence factors produced by planktonic and biofilm cell forms of this pathogen was assessed. Significant enhancement in growth and elaboration of all the virulence determinants by both the cell forms was observed when P. aeruginosa was grown in presence of supernatants with macrophage secretory products. Implications of these findings in relation to urinary tract infections induced by P. aeruginosa have been discussed.     

Effect of long-term application of rice straw,farmyard manure and inorganic fertilizer on potassium dynamics in soil

Knowledge of K-dynamics in soils can help devise practices for efficient K management in intensive rice-wheat systems. We studied the effect of long-term application of rice straw, farmyard manure (FYM) and inorganic fertilizer on total K and its distribution among different forms in 60-cm soil profile after 14 years of rice-wheat cropping. The exchangeable, the non-exchangeable and the lattice K respectively comprised 1%, 3–10% and 89–95% of total K in surface soil under different treatments. Application of rice straw and FYM positively impacted total K status of soil and its distribution among different forms. The greatest concentrations of total K, lattice K, exchangeable K and NH₄OAc-extractable K were observed in plots receiving both rice straw and FYM together and the lowest in inorganic fertilizer treated plots. On the contrary, the non-exchangeable K was the highest in inorganically fertilized plots and the lowest in rice straw amended plots. The exchangeable, the water soluble and the NH₄OAc-extractable K decreased with soil depth and did not indicate K movement beyond the rooting zone of the crops. The results showed that incorporation of rice residue in soil, instead of burning, besides reducing environmental pollution led to improved K-fertility of soils.     

New generation of protox-inhibiting herbicides

Benzfendizone belongs to the 2-[(4-heterocyclic-phenoxymethyl)-phenoxy]alkanoate class of herbicides which are potent inhibitors of the plant enzyme protoporphyrinogen oxidase (Protox). Benzfendizone, when applied postemergence, provides control of a number of key grass and broadleaf weeds in orchards and no-till crop situations. Benzfendizone can also be used as a potato desiccant and cotton defoliant. Crops such as soyabean and corn showed no injury when planted 2–3 days following postemergence application of benzfendizone.     

Cryopreservation of isolated blastomeres and embryonic stem‐like cells of Leopard danio,Brachydanio frankei

This study aimed at developing a suitable cryopreservation protocol for embryonic stem (ES)‐like cells of a tiny freshwater fish Leopard danio (Brachydanio frankei). Embryonic stem (ES)‐like cells derived from blastomeres of the early blastulae stage of the developing embryo were cultured in vitro in a medium containing Leibowitz‐15 supplemented with 10% foetal bovine serum, leopard danio embryo extract, sodium bicarbonate, sodium selenite, basic fibroblast growth factor, epidermal growth factor and leukaemia inhibitory factor. The ES‐like cells showed properties similar to ES cells in other species. They were morphologically small, round to polygonal and present in patches and extensively expressed alkaline phosphatase and stage‐specific embryonic antigen. The toxicity and chilling sensitivity of these cells were determined using ethylene glycol (EG), propylene glycol (PG) and glycerol as cryoprotective agents at molar concentrations of 0.6, 1.0, 1.4, 1.8 and 2.0. Among them, 1.8 M EG showed 70% significant viable ES‐like cells (P<0.05). The post‐thawed cells retained similar properties of non‐cryopreserved ES‐like cells with a viability rate of 65%. Similarly, blastomeres cryopreserved following the slow cooling rate with EG and PG yielded a viability of more than 70%.     

Prevalence and risk factors associated with Cryptosporidium spp. infection in young domestic livestock in India

A total of 938 faecal samples (461 cattle calves, 264 buffalo calves, 55 lambs, 116 kids and 42 piglets) from different livestock farms and individual small holdings in six targeted states of India were collected and screened by modified Ziehl–Neelsen staining technique to determine the prevalence of Cryptosporidium spp. and its association with age, sex, season and faecal consistency in domesticated animals. Overall, 16.2 % of the animals were positive for Cryptosporidium infection with prevalence of 16.3, 24.2, 1.8, 3.5 and 19.1 % in cattle calves, buffalo calves, lambs, kids and piglets, respectively. The prevalence of infection was significantly higher (p?<?0.05) in bovines (19.3 % cattle and 33.7 % buffalo) below 1 month of age than in animals between 1 and 3 months of age. But in piglets, it was higher in the age group of 1 to 3 months (22.6 %) than in younger animals (9.1 %). Also, higher prevalence (p?>?0.05) was recorded in females than in males. Seasons had a significant effect (p?<?0.05) on the prevalence of infection in large ruminants, with the highest prevalence in monsoon (cattle 28.8 % and buffalo 36.6 %) followed by pre-monsoon and post-monsoon season. However, in case of sheep and goats, the prevalence was higher (p?>?0.05) in post-monsoon than in monsoon season. A high degree of association was noticed between Cryptosporidium infection and diarrhoea in ruminants screened during the present study. But, in case of pigs, the prevalence was higher in non-diarrhoeic than in diarrhoeic animals. Genotyping of Cryptosporidium spp. based on nested PCR amplification of partial 18S rRNA and its subsequent digestion with SspI, VspI and MboII restriction enzymes revealed prevalence of Cryptosporidium parvum in representative number of positive samples of cattle, buffalo and goats.     

Regeneration ecology and population status of a critically endangered and endemic tree species （Ilex khasiana Purk.） in north-eastern India

Detailed studies on population structure and regeneration ecology of Ilex khasiana were carried out at five representative natural populations i.e. Mylliem (Population - I), Upper Shillong (Population - II), Shillong Peak (Population - III), Laitkor (Population - IV) and Nongpiyur (Population - V) to probe into the responsible affecting factors for its population structure and regeneration ability in these populations. Experimental observation under three controlled light conditions (full sunlight (100%), intermediate sunlight (50%) and low sunlight (30%)) showed that the growth status and survival rates of seedlings under intermediate and low light were better than those under the high light condition. The density-diameter distribution in population of adult trees (≥5cm dbh) exhibited a typical character of a regenerating population. A higher seedling mortality rate in natural populations of I. khasiana was most probably due to increase in light intensity following vegetation destruction and other anthropogenic disturbances that cause opening of forest canopy. The findings of the present study would be of immense value in formulating appropriate conservation measures for the species.     

An optimised protocol for molecular identification of Eimeria from chickens

Molecular approaches supporting identification of Eimeria parasites infecting chickens have been available for more than 20 years, although they have largely failed to replace traditional measures such as microscopy and pathology. Limitations of microscopy-led diagnostics, including a requirement for specialist parasitological expertise and low sample throughput, are yet to be outweighed by the difficulties associated with accessing genomic DNA from environmental Eimeria samples. A key step towards the use of Eimeria species-specific PCR as a sensitive and reproducible discriminatory tool for use in the field is the production of a standardised protocol that includes sample collection and DNA template preparation, as well as primer selection from the numerous PCR assays now published. Such a protocol will facilitate development of valuable epidemiological datasets which may be easily compared between studies and laboratories. The outcome of an optimisation process undertaken in laboratories in India and the UK is described here, identifying four steps. First, samples were collected into a 2% (w/v) potassium dichromate solution. Second, oocysts were enriched by flotation in saturated saline. Third, genomic DNA was extracted using a QIAamp DNA Stool mini kit protocol including a mechanical homogenisation step. Finally, nested PCR was carried out using previously published primers targeting the internal transcribed spacer region 1 (ITS-1). Alternative methods tested included sample processing in the presence of faecal material, DNA extraction using a traditional phenol/chloroform protocol, the use of SCAR multiplex PCR (one tube and two tube versions) and speciation using the morphometric tool COCCIMORPH for the first time with field samples.