Markers/parameters for the evaluation of natural resistance status of small ruminants against gastrointestinal nematodes

The high prevalence of anthelmintic-resistant gastrointestinal nematodes (GINs) throughout the world has led to the need for alternative worm control strategies. One of the possible substitutes to reduce the problems of drug resistance and residue is the evaluation/breeding of small ruminants for greater resistance to the GINs (organically produced), which in turn would be a helpful tool to predict the performance of an animal. At present, the existing diversity in the genetic potential to resist/tolerate GINs infection both within and between breeds has been validated. Successful selection of animals to define the genotype and identified resistance is related to the employed markers. A number of phenotypic traits such as faecal egg count (FEC), worm burden, serum antibodies, peripheral eosinophilia, packed cell volume, live weight, serum protein and albumin concentrations have been used for this purpose both in natural and artificial infections. Relatively resistant/tolerant animals have also been found to have mastocytosis, globule leucocytes, high levels of histamine and immunoglobulin (Ig) A and IgE concentrations. Of these traits, the principal and most practical measurement used to assess resistance status in animals undergoing similar parasite challenges is FEC. FEC has a positive/negative correlation with other biochemical, cellular and immunological parameters; however, the reliability of individual trial is often questioned and valuable information regarding the genetic makeup can be obtained from pooled data of a large number of trials and parameters. This paper covers all the aspects reported in the literature on various parameters considered to evaluate the resistance status of a range of small ruminant breeds.     

Enhancing protein crystallization through precipitant synergy

Suitable conditions for protein crystallization are commonly identified by screening combinations of independent factors that affect crystal formation. Because precipitating agents are prime determinants of crystallization, we investigated whether a systematic exploration of combinations of mechanistically distinct precipitants would enhance crystallization. A crystallization screen containing 64 precipitant mixtures was devised. Tests with ten HIV envelope-related proteins demonstrated that use of precipitant mixtures significantly enhanced both the probability of crystallization as well as the quality of optimized crystals. Tests with hen egg white lysozyme generated a novel C2 crystal from a salt/organic solvent mixture; structure solution at 2 A resolution revealed a lattice held together by both hydrophobic and electrostatic dyad interactions. The results indicate that mechanistically distinct precipitants can synergize, with precipitant combinations adding unique dimensions to protein crystallization.     

Cobinamides as structural probes in B12-biosynthesis: synthesis and spectroscopic analysis of isomers of Co alpha,Co beta-dicyanocobinamide

Vitamin B(12) and its coenzyme forms are cobalamins (i.e., cobamides, 'complete' with a 5,6-dimethylbenzimidazole nucleotide base), in which the particular corrinoid moiety of the cobinamides is conjugated to alpha-ribazole-3'-phosphate via a phosphate-diester group. Aside of being provided with their particular reactivity, required for their functions as organometallic cofactors in B(12)-dependent enzymes, the cobalamins also depend upon their specific three-dimensional buildup, to be able to adapt the unique constitution of 'base-on' corrinoids by intramolecular Co-coordination of the nucleotide base. We report rational partial syntheses and detailed spectral analyses of three close cobinamide isomers in their Co(alpha),Co(beta)-dicyano forms: of 13-epicobinamide (also called neocobinamide), of 176(S)-epicobinamide, and of 176-isocobinamide. Neocobinamide was obtained under acidic conditions as a degradation product of vitamin B(12). 176(S)-Epicobinamide and 176-isocobinamide were prepared by condensation of cobyric acid with (2S)-1-aminopropan-2-ol and with 3-aminopropan-1-ol, respectively. Natural cobinamide represents the corrinoid nucleus produced by proper microbial biosynthesis (as intermediate for the further assembly of the 'complete' corrinoid cofactors) or is required in some microorganisms, such as Escherichia coli, as an exogenously supplied unit for further biosynthetic buildup. The three compounds may thus be of use as structural probes for the biosynthetic capacity and tolerance in microorganisms, and (some of them) may serve as substrates as well, for further biosynthetic 'completion' of corrinoid cofactors or their analogues.     

Genetic Testing of Non-familial Deaf Patients for <Emphasis Type="Italic">CIB2</Emphasis> and <Emphasis Type="Italic">GJB2</Emphasis> Mutations: Phenotype and Genetic Counselling

CIB2 and GJB2 genes variants contribute significantly in familial cases of prelingual recessive hearing loss (HL). This study was aimed to determine the CIB2 and GJB2 variants and associated phenotype in 150 non-familial individuals with HL. After getting informed consent, 150 non-familial deaf patients were enrolled and blood samples were obtained for DNA extraction. Pure tone air conduction audiometry was performed. Coding exons of CIB2 and GJB2 genes were Sanger sequenced. A tetra primer ARMS assay was developed for recurrent CIB2 variant. Four bi-allelic GJB2 variants, c.71G>A p.(Trp24*), c.231G>A p.(Trp77*), c.235delC p.(Leu79Cysfs3*) and c.35delG p.(Gly11Leufs24*), were found in nine hearing impaired individuals. We also found four homozygotes and five carriers of c.380G>A p. (Arg127His) variant of controversial clinical significance. CIB2 sequencing revealed single recurrent variant c.272T>C p. (Phe91Ser) segregating with HL in ten individuals. Among our patients, c.71G>A (p.Trp24*) was the most common variant, accounted for 45% of GJB2 variants. Two known GJB2 variants, c.235delC p. (Leu79Cysfs3*) and c.310del14 p. (Lys105Argfs2*), are reported here for the first time in Pakistani population. Our data further support the benign nature of c.380G>A p. (Arg127His) variant. For CIB2, c.272T>C p. (Phe91Ser) is the second common cause of HL among our sporadic cases. Phenotypically, in our patients, individuals homozygous for GJB2 variants had profound HL, whereas CIB2 homozygotes had severe to profound prelingual HL. Our results suggest that GJB2 and CIB2 are common cause of HL in different Pakistani ethnicities.     

Endophytic bacterial diversity of Avicennia marina helps to confer resistance against salinity stress in Solanum lycopersicum

The current study aimed to explore the endophytic bacterial diversity of Avicennia marina and the potential roles of these endophytes in counteracting saline conditions in tomato plants. Molecular analysis revealed strains from Paenibacillus, Bacillus, Microbacterium, Citrobacter, Lysinibacillus, Halomonas, Virgibacillus, Exiguobacterium, and Vibrio. However, Bacillus pumilus AM11 and Exiguobacterium sp. AM25 showed significantly higher growth in saline media. In response to salinity stress, tomato plants treated with AM11 and AM25 showed significantly higher (～15–23%) biomass, photosynthetic rate and pigment accumulation compared to controls. Salinity-exposed plants had significantly reduced growth and increased (three-fold) lipid peroxidation, whilst glutathione, catalase, and peroxidase activities were significantly reduced. In contrast, AM11, AM25, and methionine improved these physiochemical attributes. The study concludes that the application of bacterial endophytes from plants growing in saline conditions can offer other plants similar stress-resistance potential. Such halophytic bacterial strains can be used to improve plant growth in saline conditions.     

Characterization of a Novel Enterovirus Serotype and an Enterovirus EV-B93 Isolated from Acute Flaccid Paralysis Patients

Non-polio enteroviruses (NPEVs) are among the most common viruses infecting humans worldwide. Most of these infections are asymptomatic but few can lead to systemic and neurological disorders like Acute Flaccid Paralysis (AFP). Acute Flaccid Paralysis is a clinical syndrome and NPEVs have been isolated frequently from the patients suffering from AFP but little is known about their causal relationship. The objective of this study was to identify and characterize the NPEV serotypes recovered from 184 stool samples collected from AFP patients in Federally Administered Tribal Areas (FATA) in north-west of Pakistan. Overall, 44 (95.6 %) isolates were successfully typed through microneutralization assay as a member of enterovirus B species including echovirus (E)-2, E-3, E-4, E-6, E-7, E-11, E-13, E-14, E-21 and E-29 while two isolates (PAK NIH SP6545B and PAK NIH SP1202B) remained untypeable. The VP1 and capsid regions analysis characterized these viruses as EV-B93 and EV-B106. Phylogenetic analysis confirmed that PAK NIH isolates had high genetic diversity and represent distinct genotypes circulating in the country. Our findings highlight the role of NPEVs in AFP cases to be thoroughly investigated especially in high disease risk areas, with limited surveillance activities and health resources.     

Serotype Diversity of Astroviruses in Rawalpindi,Pakistan during 2009–2010

Astroviruses are globally known enteropathogens causing gastroenteritis and diarrhea, with eight well defined serotypes. Epidemiological studies have recognized serotype-1 as the most common subtype but no such data is available in Pakistan. During 2009–2010, we found astroviruses in 41 out of 535 (7%) samples collected from hospitalized children. Thirty one strains belonged to serotype-1 and clustered into two distinct lineages. Serotype-3, -4 and -6 were detected with 97–98% genetic homology to Indian and Chinese strains.     

Epidemiology and Genetic Diversity of Rotavirus Strains in Children with Acute Gastroenteritis in Lahore,Pakistan

Pakistan harbors high disease burden of gastro-enteric infections with majority of these caused by rotavirus. Unfortunately, lack of proper surveillance programs and laboratory facilities have resulted in scarcity of available data on rotavirus associated disease burden and epidemiological information in the country. We investigated 1306 stool samples collected over two years (2008–2009) from hospitalized children under 5 years of age for the presence of rotavirus strains and its genotypic diversity in Lahore. The prevalence rate during 2008 and 2009 was found to be 34% (n = 447 out of 1306). No significant difference was found between different age groups positive for rotavirus (p>0.05). A subset of EIA positive samples was further screened for rotavirus RNA through RT-PCR and 44 (49.43%) samples, out of total 89 EIA positive samples, were found positive. G and P type prevalence was found as follows: G1P [4] = 3(6.81%); G1P [6] = 9(20.45%); G1P [8] = 1(2.27%); G2P [4] = 21(47.72%); G2P [8] = 1(2.27%); G9P [4] = 1(2.27%); G9P [6] = 1(2.27%) and G9P [8] = 7(15.90%). Phylogenetic analysis revealed that the VP7 and VP4 sequences clustered closely with the previously detected strains in the country as well as Belgian rotaviruses. Antigenic characterization was performed by analyzing major epitopes in the immunodominant VP7 and VP4 gene segments. Although the neutralization conferring motifs were found variable between the Pakistani strains and the two recommended vaccines strains (Rotarix™ and RotaTeq™), we validate the use of rotavirus vaccine in Pakistan based on the proven and recognized vaccine efficacy across the globe. Our findings constitute the first report on rotavirus’ genotype diversity, their phylogenetic relatedness and epidemiology during the pre-vaccination era in Lahore, Pakistan and support the immediate introduction of rotavirus vaccine in the routine immunization program of the country.     

Preussia sp. BSL-10 producing nitric oxide,gibberellins, and indole acetic acid and improving rice plant growth

Preussia sp. have been least known to improve plant growth and produce phytohormones. The current study investigated the production of nitric oxide (NO), indole-3-acetic acid (IAA), and gibberellins (GA₄, GA₇, GA₁₅, and GA₅₃) by a novel endophytic-fungal strain Preussia sp. BSL-10 using advanced chromatographic and spectroscopic techniques. Production of these phytohormones were validated by RT–PCR analysis, which indicated the expression of genes encoding tryptophan synthase (TRP), indole-3-acetamide hydrolase (IAAH), tryptophan-2-monooxygenase (IAAM), aldehyde dehydrogenase (ALD), GA₄ desaturase (DES), geranylgeranyl-diphosphate synthase (GGS2), ent-desaturase oxidase (P450-4), GA₁₄ synthase (P450-1) and nitrite reductase (NIRK/NIRS), cytochrome P450 (P450nor), nitrate reductase (NR), NOS-like (NOL), and nitric oxide reductase (QNOR/CNOR). In plant growth-promoting effects, the inoculation of Preussia sp. BSL-10 significantly increased the growth of dwarf mutant Waito-C and wild-type rice cultivars. In conclusion, utilizing new endophytic with the ability to produce NO, IAA, and gibberellins can be used to promote growth and yield of marginalized crops.     

Ethylene Supplementation Increases PSII Efficiency and Alleviates Chromium-Inhibited Photosynthesis Through Increased Nitrogen and Sulfur Assimilation in Mustard

Journal of Plant Growth Regulation - The effect of chromium (Cr) toxicity on four cultivars of mustard (Brassica juncea), Varuna, Pusa Bold, Rohini and SS2, was studied to select the cultivar with...