Climate variability is highly impacting on mosquito-borne diseases causing malaria and dengue fever across the globe. Seasonal variability change in temperature and rainfall patterns are impacting on human health. Mosquitoes cause diseases like dengue fever, yellow fever, malaria, Chikungunya, West Nile and Japanese encephalitis. According to estimations by health organizations, annually one million human deaths are caused by vector-borne diseases, and dengue fever has increased about 30-fold over the past 50 years. Similarly, over 200 million cases of malaria are being reported annually. Mosquito-borne diseases are sensitive to temperature, humidity and seasonal variability. Both conventional (environmental, chemical, mechanical, biological etc.) and nanotechnology-based (Liposomes, nano-suspensions and polymer-based nanoparticles) approaches are used for the eradication of Malaria and dengue fever. Now green approaches are used to eradicate mosquitoes to save human health without harming the environment. In this review, the impact of climatic conditions on mosquito-borne diseases along with conventional and nanotechnology-based approaches used for controlling malaria and dengue fever have been discussed. Important recommendations have been made for people to stay healthy.

Red blotch is an emerging disease of grapevine associated with grapevine red blotch-associated virus (GRBaV). The virus spreads with infected planting stocks but no vector of epidemiological significance has been conclusively identified. A vineyard block of red-blotch-affected Vitis vinifera ‘Cabernet franc’ clone 214 was observed in California, with a clustering of infected, symptomatic vines focused along one edge of the field proximal to a riparian habitat with free-living Vitis spp. No genetic heterogeneity was observed in a 587-nucleotide region of the GRBaV genome in a population of 44 Cabernet franc clone 214 isolates. By contrast, genetic differences were observed in isolates from other cultivars and clones growing in adjacent blocks. GRBaV was confirmed infecting four free-living vines, two of which were shown to be V. californica × V. vinifera hybrids. The genomes of three free-living GRBaV vine isolates and seven from V. vinifera cultivars were compared; free-living vine isolates were shown to be more similar to each other and a ‘Merlot’ isolate than to the other cultivated vine isolates. The finding that GRBaV is present in free-living Vitis spp. indicates the virus can be spread by natural (nonhuman-mediated) means, and we hypothesize that in-field spread of GRBaV is occurring.

Human Papillomavirus (HPV) is well known pathogen that can cause benign and malignant tumors in humans, yet there is very little information regarding HPV types prevalent in Pakistan. A total of 92 cervical secretions were collected from suspected married female patients and used for DNA isolation using a novel isolation method. The samples were tested through Polymerase Chain Reaction (PCR) using already reported primers MY09/MY11, GP5/GP6, GP5+/GP6+, CP65/CP70, CP66/CP69 and SPF1/SPF2 and with those developed in this study including HRT1 and HRT2 primer sets for typing HPV types and HACTB primer set for human beta actin gene as internal positive control. Sequencing and phylogenetic analyses were performed for two isolates to determine circulating HPV types. PCR with HRT1 and HRT2 indicated 2 (2.17 %) patients were positive for HPV type- 16 while 1 (1.08 %) with HPV type 18. Sequencing and phylogenetic analysis of isolates confirmed HPV type-16 in genus alpha 9 which have 99 % homology with already reported HPV from Japan and Costa Rica. This is the first report of HPV type-16 genus alpha 9 in Pakistan and the reported assay and sequence data will serve as valuable tools in further epidemiological studies for HPV surveillance to improve public health, especially of females in Pakistan.

Germin and germin-like proteins (GLPs) are water soluble extracellular proteins reportedly expressed in response to some environmental and developmental signals. Some enzymatic activities have also been associated with germin/GLPs. However, their role in overall metabolism has not been fully understood. Significant insight into their function may also be gained by analysis of their promoter. During this study, about 1107 bp 5′region of OsRGLP2 gene was amplified, cloned and sequenced. The sequence analysis by BLAST showed that this promoter sequence has five common regions (CR1–CR5) of different sizes, which are repeated at 3–6 other locations in 30 kb region in which this gene driven by its promoter is located. Interestingly, all the genes driven by promoter harboring these common regions are GLPs/putative germins. Analysis of these common regions located on OsRGLP2 indicated presence of many elements including those for light responsiveness, dehydration and dark induced senescence, stresses (pathogen and salt), plant growth regulators, pollen specific expression and elements related to seed storage proteins. Analysis of the 30 kb germin/GLP clustered region by GenScan detected each gene to have a putative 40 bp promoter which contains TATA box and Dof factor which turned out to be a part of CR2.

The success of Agrobacterium mediated plant transformation depends to a certain extent on appropriate selection of the A. tumefaciens strain for a particular plant species. Many stages in a plant transformation procedure are prone to bacterial contamination with similar antibiotic resistance that may compromise the identity of the A. tumefaciens strain used, in turn adversely affecting success of a transformation experiment. Different primer sets were designed to exploit genetic differences among different strains of A. tumefaciens which are commonly used for plant genetic transformation, to identity confirmation as well as to distinguish them from one another. The primer sets Ach5FtsZ-F/R specific for Ach5 and C58GlyA-F/R specific for C58 were designed on chromosomal DNA while primer sets pTiBo542-F/R and nptI-F/R specific for plasmid pTiBo542 are capable to identify and distinguish these strains from one another. These primer sets when used simultaneously in multiplex PCR, produce a pattern which uniquely identifies all these strains and distinguishes them except for GV3101 and C58C1, which can further be distinguished from each other by rifampicin screening. The multiplex PCR assay and primers being reported here serve as a valuable tool in determining the identity of A. tumefaciens strains at any stage of plant transformation procedure.

Banana bunchy top disease (BBTD), caused by a Banana Bunchy Top Virus (BBTV), is the most devastating viral disease of bananas. The genome of multipartite BBTV is about 1 Kb in size and comprises six circular single-stranded (ss) DNA components, namely DNA-R, C, S, M, U3, and N. Gene expression in multipartite DNA viruses is highly flexible due to variations in the copy number and transcript levels of individual genomic components. This enables them to adapt to changing conditions and maintain optimal fitness. In the current study, the copy number of each genetic component of BBTV and its transcript level were determined in the leaf, midrib, and root tissue of the infected banana plant. The results of the study showed that DNA copies (copies/ng) were higher in midrib (11,105 ± 10,704.42), followed by leaf (6902.91 ± 1651.14), and were lowest in root (23.49 ± 9.21) tissues. In contrast, the transcript level was highest in the root (312.45 ± 106.69) and lowest in the midrib (0.72 ± 0.16). This suggests that BBTV independently regulates the transcriptional level of each DNA component. The distribution of BBTV in different tissues assists the identification of significantly higher viral load tissues for early and sensitive disease diagnosis, which will be beneficial for better disease management.