13C nuclear magnetic resonance spectra of diastereomeric C-24 alkyl sterols have been assigned. Differences in the chemical shifts of side-chain carbons permitted the determination of the absolute configuration at C-24 in several sterols since these chemical shifts are insensitive to structural changes remote from the asymmetric centre. An unknown sterol from Tetraselmissuecica has been identified as (24R)-24-methylcholest-5-en-3β-ol and the configuration assigned from 1H nmr data to the sterol from Phaeodoctylumtricornutum has been confirmed. The utility and potential of this method in characterising new sterols and their biological precursors is discussed.

BackgroundEquine piroplasmosis (EP) caused by Theileria equi, Babesia caballi, or both, contributes to significant economic loss in the equine industry and remains uncontrolled in Egypt. This study focuses on surveying T. equi and B. caballi infections and hematological disorders in equine populations in Egypt.MethodsTheileria equi and B. caballi infections were assessed in blood from 88 horses and 51 donkeys in Egypt using light microscopy, indirect immunofluorescent antibody test (IFAT), nested PCR (nPCR), and competitive-ELISA (cELISA) assays. PCR products were examined for specificity by DNA sequencing. Hematological alterations were evaluated using a standard cell counter.ResultsMicroscopic analysis revealed EP infection in 11.4 % and 17.8 % of horses and donkeys respectively. IFAT detected 23.9 % and 17.0 % infection of T. equi and B. caballi, respectively, in horses, and 31.4 % of T. equi and B. caballi in donkeys. T. equi cELISA detected 14.8 % and 23.5 % positive horses and donkeys, respectively, but the B. caballi RAP-1-based cELISA failed to detect any positives, a result hypothesized to be caused by sequence polymorphism found in the rap-1 genes. Nested-PCR analysis identified 36.4 % and 43.1 % positive horses and donkeys, respectively for T. equi and it also identified 19.3 % and 15.7 % positive horses and donkeys, respectively for B. caballi. The overall EP incidence found in the population under study was relatively high and comparable regardless of the diagnostic method used (56.8 % using nPCR and 48.9 % using IFAT). Hematologic analysis revealed macrocytic hypochromic anemia and thrombocytopenia in all piroplasma-infected horses.ConclusionsThe data confirm relatively high levels of EP, likely causing hematological abnormalities in equines in Egypt, and also suggest the need for an improved serological test to diagnose B. caballi infection in this region.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1539-9) contains supplementary material, which is available to authorized users.

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Aim:This work aimed to study epidemiology and diagnosis of feline panleukopenia virus (FPV) using clinical examination, direct ELISA, RNA viral isolation and identification, and knowing phylogenetic tree of our isolate.Materials and Methods:One hundred and sixty-five cats of different ages and sex were examined. Each cat was examined clinically to detect the clinical manifestations of the disease showing symptoms suggestive of feline panleukopenia (FP) as well as ELISA, and polymerase chain reaction (PCR) amplification analyses were conducted.Results:Our finding includes (a) clinical signs detected in 165 of 165 cats were in the form of lethargy, fever, anorexia, thirst, vomiting, diarrhea, dehydration, and leukopenia. (b) ELISA results revealed that 66 of all examined cats were positive for FPV. (c) The amplification products from all positive samples were confirmed as FPV (VP1) gene by nucleotide sequences analysis, in which 75 samples were positive using PCR amplification for the FPV. (d) Statistical evaluation of ELISA results in comparison to PCR findings. ELISA showed 88%, 100%, and 94.5% for sensitivity, specificity, and accuracy, respectively, while the prevalence of FP among the examined population was 45%. No effect of sex, breed, and age on ELISA results as recorded using Chi-square analysis.Conclusion:The results of the sequence analysis indicated that PCR products of the FPV cDNA exhibited very low variation in their nucleotide sequence of all isolates compared with the published FPV genome, which could be suggested that FPV appears to be genomically stasis compared with other Parvoviruses. The genome sequence of FPLV strain in this study has been deposited in GenBank under the accession number KY466003. Our isolate closely related 100% to isolates from Portugal, which might be the origin of infection to Egypt through importation of cats.

Ethylenediaminetetraacetic acid (EDTA) is widely used in food and other industries to sequester metal ions and to prevent their disadvantageous effects. The objective of the current study was to evaluate the protective effect of Panax ginseng extract standardized with ginsenoside Rg3 (ginsenoside Rg3 content was 3.6% w/w, i.e., 36 microg/mg P. ginseng extract) and garlic against EDTA-induced biochemical, genotoxic, and histological changes in rats. Forty male rats were divided into eight treatment groups and treated for 7 days as follows: the control group, the group treated with EDTA (20 mg/kg b.w) and the groups treated with P. ginseng extract (20 mg/kg b.w), garlic (5 mg/kg b.w), P. ginseng plus garlic alone or in combination with EDTA. In vivo bone marrow micronucleus test and random amplified polymorphism DNA-PCR (RAPD-PCR) method were performed to assess the antigenotoxic effect of both protective agents. The results indicated that EDTA administration caused a significant decrease in the serum biochemical parameters and antioxidant enzymes activity. The administration also increased lipid peroxidation and the incidence of micronucleated polychromatic erythrocytes (MnPCEs), caused appearance of some changes in polymorphism band patterns, and induced different histopathological lesions in the livers, kidneys, and testis. Treatment with P. ginseng, garlic alone or plus EDTA significantly improved all the tested parameters. Moreover, P. ginseng extract was found to be more effective than garlic in restoring the parameters that were altered by EDTA.