Biochemical characterization of Taenia hydatigena cysticerci from goats and pigs

Analysis of the major biochemical components of Taenia hydatigena cysticerci collected from goats and pigs showed marked differences, particularly in glycogen, protein, lipid and DNA levels. Differences were also detected in the levels of cholesterol, triglycerides, free fatty acids and phospholipids. Furthermore, the profile of phospholipid fractions revealed quantitative differences between the two species. It is concluded that the cysticerci of goat and pig origin probably represent two different strains and possibly follow the same pattern of speciation as reported in the related taeniid, Echinococcus granulosus.     

“Occurrence of blaCTX-MGp1 and blaCTX-MGp26 in third generation cephalosporin-resistant and carbapenem- resistant bacterial isolates from southwest region of Saudi Arabia-a preliminary study”

This study was intended to identify the genes responsible for ESBL- and carbapenemase-producing bacterial isolates obtained from Jizan region. A hospital-based cross-sectional study was conducted over a period of 3 months (15th November 2018–15th February 2019). Fifty non-duplicate, 3rd-generation cephalosporin and carbapenem-resistant isolates were collected from microbiology lab of a tertiary care hospital in Jizan province and were screened for ESBLs and MBLs by phenotypic methods (CDT). The positive isolates (by phenotypic method) were then scanned for the presence of bla_ESBLs and bla_NDM-₁ genes, respectively, by PCR.As a result, 10% isolates showed imipenem-cephalosporin co-resistance whereas 92% (46/50) of isolates were found to be ESBL producers by CDT. The maximum occurrence was observed for bla_CTX-M (70%), followed by bla_SHV (16%) and least occurrence was noted for bla_TEM (12%). Moreover, 97% isolates (34/35) were of bla_CTX-MGroup1 but one isolate showed the presence of bla_CTX-M Group26. Despite the co-resistance of cephalosporin and carbapenem, 14% (7/50) were found to be MBL producer on phenotypic detection by Combination Disc Test (CDT), whereas all the isolates were found to be negative for bla_NDM-1. Hence bla_CTX-MGroup1 is present in quite high fraction followed by bla_SHV in the bacterial isolates of Jizan region. Moreover, the occurrence of bla_CTX-M Group1 and bla_CTX-M Group26 in clinical isolates from the Jizan region of Saudi Arabia has been reported for the first time.     

Evaluation of Pakistan wheat germplasms for stripe rust resistance using molecular markers

Wheat production in Pakistan is seriously constrained due to rust diseases and stripe rust (yellow) caused by Puccinia striiformis f. sp. tritici, which could limit yields. Thus development and cultivation of genetically diverse and resistant varieties is the most sustainable solution to overcome these diseases. The first objective of the present study was to evaluate 100 Pakistan wheat cultivars that have been grown over the past 60 years. These cultivars were inoculated at the seedling stage with two virulent stripe rust isolates from the United States and two from Pakistan. None of the wheat cultivars were resistant to all tested stripe rust isolates, and 16% of cultivars were susceptible to the four isolates at the seedling stage. The data indicated that none of the Pakistan wheat cultivars contained either Yr5 or Yr15 genes that were considered to be effective against most P. striiformis f. sp. tritici isolates from around the world. Several Pakistan wheat cultivars may have gene Yr10, which is effective against isolate PST-127 but ineffective against PST-116. It is also possible that these cultivars may have other previously unidentified genes or gene combinations. The second objective was to evaluate the 100 Pakistan wheat cultivars for stripe rust resistance during natural epidemics in Pakistan and Washington State, USA. It was found that a higher frequency of resistance was present under field conditions compared with greenhouse conditions. Thirty genotypes (30% of germplasms) were found to have a potentially high temperature adult plant (HTAP) resistance. The third objective was to determine the genetic diversity in Pakistan wheat germplasms using molecular markers. This study was based on DNA fingerprinting using resistance gene analog polymorphism (RGAP) marker analysis. The highest polymorphism detected with RGAP primer pairs was 40%, 50% and 57% with a mean polymorphism of 36%. A total of 22 RGAP markers were obtained in this study. RGAP, simple sequence repeat (SSR) and sequence tagged site (STS) markers were used to determine the presence and absence of some important stripe rust resistance genes, such as Yr5, Yr8, Yr9, Yr15 and Yr18. Of the 60 cultivars analyzed, 17% of cultivars showed a RGAP marker band for Yr9 and 12% of cultivars exhibited the Yr18 marker band. No marker band was detected for Yr5, Yr8 and Yr15, indicating a likely absence of these genes in the tested Pakistan wheat cultivars. Cluster analysis based on molecular and stripe rust reaction data is useful in identifying considerable genetic diversity among Pakistan wheat cultivars. The resistant germplasms identified with 22 RGAP markers and from the resistance evaluations should be useful in developing new wheat cultivars with stripe rust resistance.     

Preliminary studies in numerical systematics of the Paramphistomoidea (Digenea) from domestic Artiodactyla of Northern India

The systematic relationships between nine taxa (species) of paramphistomes belonging to the families Cladorchiidae, Gastrodiscidae, Gastrothylacidae and Paramphistomidae from domestic buffaloes Bubalus bubalis and pigs Sus scrofa were examined using SEM revealed two types of papillae occurring commonly on the oral cone and less frequently around the gonopore and acetabulum. Non-pitted papillae occur on the oral cone in Paramphistomum epiclitum, Calicophoron calicophorum, Explanatum (=Gigantocotyle) explanatum, Orthocoelium scoliocoelium, Gastrothylax crumenifer and Olveria indica. These papillae are also evident around the gonopore in Calicophoron papillosum, O. scoliocoelium and O. indica, and the ventral pouch opening in G. crumenifer. Ciliated papillae occur on the oral cone in C. papillosum and Fischoederius elongatus. Morphological and immunological observations were employed to classify paramphistomes using phenetic and phylogenetic approaches to systematics. Classifications based on phenetic methods compared taxa according to overall similarity of their characteristics and produced largely discordant results. Classifications based on phylogenetic methods compared taxa according to the genealogical descent of their characteristics and produced largely concordant results. These are in agreement with the widely accepted scheme of classification proposed by Stiles & Goldberger (1910) in which Gastrothylacidae and Paramphistomidae occur as natural groups. Furthermore, the Gastrothylacidae appear to share common ancestry with a member of the Paramphistomidae and are consequently nested within the latter group.This study forms part of the requirements for a thesis submitted by the first author to Q.U.B. for the degree of Doctor of Philosophy.This study forms part of the requirements for a thesis submitted by the first author to Q.U.B. for the degree of Doctor of Philosophy.     

Quantitative studies on acetylcholinesterase in seven species of digenetic trematodes

Quantitative estimation of absolute levels and in vitro release of acetylcholinesterase (AChE) in seven species of digenetic trematodes: Isoparorchis hypselobagri from the swim bladder of catfish, Wallago attu; Srivastavaia indica and Gastrothylax crumenifer from the rumen, and Gigantocotyle explanatum from the liver of the water buffalo, Bubalus bubalis; Fasciolopsis buski, Echinostoma malayanum from the small intestine and Gastrodiscoides hominis from the caecum of the pig, Sus scrofa revealed that the enzyme is present in remarkably high quantities in species which inhibit gastrointestinal tract compared with those that parasitize liver and swim bladder. The rate of in vitro release of AChE also varies with the species which supports the view that such differential secretion probably takes place in situ as well to counteract peristalsis and it is a biochemical adaptation on the part of these trematodes.     

Regulating the fate of stem cells for regenerating the intervertebral disc degeneration

Lower back pain is a leading cause of disability and is one of the reasons for the substantial socioeconomic burden. The etiology of intervertebral disc (IVD) degeneration is complicated, and its mechanism is still not completely understood. Factors such as aging, systemic inflammation, biochemical mediators, toxic environmental factors, physical injuries, and genetic factors are involved in the progression of its pathophysiology. Currently, no therapy for restoring degenerated IVD is available except pain management, reduced physical activities, and surgical intervention. Therefore, it is imperative to establish regenerative medicine-based approaches to heal and repair the injured disc, repopulate the cell types to retain water content, synthesize extracellular matrix, and strengthen the disc to restore normal spine flexion. Cellular therapy has gained attention for IVD management as an alternative therapeutic option. In this review, we present an overview of the anatomical and molecular structure and the surrounding pathophysiology of the IVD. Modern therapeutic approaches, including proteins and growth factors, cellular and gene therapy, and cell fate regulators are reviewed. Similarly, small molecules that modulate the fate of stem cells for their differentiation into chondrocytes and notochordal cell types are highlighted.     

Partial purification and characterization of Gigantocotyle explanatum somatic antigens

Soluble extracts of Gigantocotyle explanatum, isolated from the liver of buffalo Bubalus bubalis were fractionated on Sephadex G-200 columns. Nine major fractions referred to as F1, F2, F3, F4, F5, F6, F7, F8 and F9 were separated. Each fraction was tested by ELISA for antigenicity using sera from G. explanatum-infected field buffaloes. Fractions F1 and F2 were highly antigenic, F3, F4, F6 and F7 were moderately antigenic and F5, F8 and F9 were poorly antigenic. Analyses by SDS-PAGE revealed that each fraction comprised several polypeptide(s) in the molecular weight range of <29 to >205 kDa. Results of Western blotting indicated that not all polypeptides which appeared in the SDS-PAGE were antigenic. The antigenic molecules of each fraction were mostly in the low molecular weight range of <14 to >94 kDa with the polypeptides in the range of >14, 14, 18, 21-25 and 34-36 kDa.     

Analysis of excretory-secretory and somatic antigens of Gastrothylax crumenifer

The excretory/secretory (ES) metabolic products released by Gastrothylax crumenifer (Trematoda: Digenea) during in vitro incubations and the somatic extract of the adult parasite were analysed using polyacrylamide gel electrophoresis (PAGE). Immunogenicity of ES and somatic extracts were evaluated by immunoblotting and ELISA using sera raised against ES and somatic antigens in rabbits. The electropherograms of ES and somatic extracts have been resolved into 38 and 41 polypeptides, respectively. The apparent molecular weights of these polypeptides range from < 29 to > 205 kDa. A total of 14 polypeptides were found to be common to both of the samples. The analysis of immunoblot results revealed 22 and 27 antigenic polypeptides in ES and somatic extracts respectively. Only 11 and 13 antigenic polypeptides were found specific to ES and tissue extract respectively. The molecular weights of these specific polypeptides were calculated to be < 14.4, 16, 20, 25, 33, 42, 119, 125 and > 205 kDa for metabolic products and < 14.4, 25, 30, 35, 78, 84 and > 205 kDa for the tissue extracts, respectively. Analysis of ELISA results revealed that a dilution of up to 1:3200 of the test sera could react with the ES product. Further, when the ES antigens were allowed to react with antisomatic extracts in hyperimmune sera the titre of IgG increased up to a dilution of 1:12800. The potential importance of these antigens in the immunodiagnosis of amphistomiasis is discussed.