Ahmed Elsify

Evaluation of in vitro and in vivo inhibitory effects of fusidic acid on Babesia and Theileria parasites

Fusidic acid known to has antibacterial, antifungal, and antimalarial activities. Fusidic acid blocks translation elongation factor G gene in Plasmodium falciparum. In the present study, the inhibitory effects of fusidic acid on the in vitro growth of bovine and equine Babesia parasites were evaluated. The inhibitory effect of fusidic acid on the in vivo growth of Babesia microti was also assessed. The in vitro growth of four Babesia species that were tested was significantly inhibited (P<0.05) by micromolar concentrations of fusidic acid (IC(50) values=144.8, 17.3, 33.3, and 56.25 μM for Babesia bovis, Babesia bigemina, Babesia caballi, and Theileria equi, respectively). Combinations of fusidic acid with diminazene aceturate synergistically potentiated its inhibitory effects in vitro on B. bovis and B. caballi. In B. microti-infected mice, fusidic acid caused significant (P<0.05) inhibition of the growth of B. microti at the dose of 500 mg/kg BW relative to control group. These results indicate that fusidic acid might be incorporated in treatment of babesiosis.

Prevalence and Identity of Taenia multiceps cysts “Coenurus cerebralis” in Sheep in Egypt

Coenurosis is a parasitic disease caused by the larval stage (Coenurus cerebralis) of the canids cestode Taenia multiceps. C. cerebralis particularly infects sheep and goats, and pose a public health concerns. The present study aimed to determine the occurrence and molecular identity of C. cerebralis infecting sheep in Egypt. Infection rate was determined by postmortem inspection of heads of the cases that showed neurological manifestations. Species identification and genetic diversity were analyzed based on PCR-sequence analysis of nuclear ITS1 and mitochondrial cytochrome oxidase (COI) and nicotinamide adenine dinucleotide dehydrogenase (ND1) gene markers. Out of 3668 animals distributed in 50 herds at localities of Ashmoun and El Sadat cities, El Menoufia Province, Egypt, 420 (11.45%) sheep showed neurological disorders. Postmortem examination of these animals after slaughter at local abattoirs indicated to occurrence of C. cerebralis cysts in the brain of 111 out of 420 (26.4%), with overall infection rate 3.03% of the involved sheep population. Molecular analysis of representative samples of coenuri at ITS1 gene marker showed extensive intra- and inter-sequence diversity due to deletions/insertions in the microsatellite regions. On contrast to the nuclear gene marker, considerably low genetic diversity was seen in the analyzed mitochondrial gene markers. Phylogenetic analysis based on COI and ND1 gene sequences indicated that the generated sequences in the present study and the reference sequences in the database clustered in 4 haplogroups, with more or less similar topologies. Clustering pattern of the phylogenetic tree showed no effect for the geographic location or the host species.

Animal level risk factors associated with Babesia and Theileria infections in cattle in Egypt

In present study, blood samples were collected randomly from 439 cows at three main regions of Egypt (northern, central and southern). Molecular diagnosis of Babesia and Theileria infections by PCR amplification of DNA (gene) fragments, then cloning and sequencing of the positive samples were conducted. A questionnaire was created to imply the assumed risk factors and logistic regression statistical analysis was carried out to appraise the potential factors on the animal level. The results revealed that 49 (11.16%) and 45 (10.25%) cattle were infected with Babesia and Theileria parasites, respectively. B. bigemina (7.97%) and T. annulata (9.56%) were the most prevalent parasites. For Babesia sp., final multivariate logistic regression analysis showed a significant association between the infection and irregular use of antiprotozoal drugs (P = 0.003; OR: 0.28; 95% CI: 0.12–0.65), management practice (P = 0.029; OR: 6.66; 95% CI: 1.21–36.59) and ecology area (P = 0.006; OR: 5.62; 95% CI: 1.63–19.31). However, for Theileria sp. infection, animal breed (P = 0.003; OR: 0.44; 95% CI:.45–1.00) and irregular use of antiprotozoal drugs (P<0.001; OR: 4.22; 95% CI: 2.62–5.60) were the potential risk factors. The results of the present study declare the prevalent bovine Babesia and Theileria sp. in Egypt based on molecular description. An impression on the potential risk factors associated with infections was obtained. Recognition of the potential risk factors associated with tick borne disease may be helpful to construct the best preventive measures.

Serum vitamin A and E, copper, zinc and selenium concentrations and their relationship with health outcomes in dromedary hospitalized camels ( Camelus dromedarius )

The goals of this study were to measure serum vitamin A (retinol) and E (α-tocopherol) and trace elements concentrations (copper, zinc and selenium) during diseases condition and to determine their association with hematological parameters and immune status of hospitalized camels. A total of 95 dromedary camels [healthy (n=65); hospitalized camels (n=30)] were included in this study. Vitamin A and E concentrations were significantly lower in hospitalized camels than apparently healthy ones (P<0.05). Hospitalized camels had lower concentrations of zinc and selenium compared to healthy camels (P<0.05). Vitamin E, copper, zinc and selenium concentrations were positively correlated with phagocytic activity in hospitalized camels (P<0.05). The likelihood of deficiency of vitamin A and E, zinc and selenium concentrations were significant in female hospitalized camels than males and in young age hospitalized camels < 6 years old compared to old ones (P<0.05). Decreased vitamin A and E and trace elements concentrations were associated with hospitalized camels’ phagocytic activity and index. The prevalence of low vitamin A and E, zinc and selenium concentrations were frequent in female hospitalized camels and hospitalized camels of age < 6 years old suggesting severe oxidative stress.

Development and characterization of ORF68 negative equine herpes virus type–1, Ab4p strain

Equine herpesvirus-1 (EHV-1) is an important pathogen, which infects horses worldwide with high morbidity but low mortality rates. The respiratory disorders and abortions are the most common indicators. Ab4p (an abortigenic and paralytic virus) is one of the most important and virulent strains. The development and functional characterization of the open reading frame-68 (ORF68) negative EHV-1 Ab4p mutants and an assessment of their roles in the infection at the cellular level were the main targets of the current study. Escherichia coli DH10β containing the Ab4p bacterial artificial chromosome (pAb4pBAC) and Red/ET expression vector were used to develop different ORF68 mutants. Multi-step growth kinetic experiments were conducted in order to evaluate the growth properties of the constructed mutant viruses. Growth of the Ab4pΔORF68 showed the lowest titer, compared to the Ab4pΔORF68R, Ab4pΔORF68R non-sense, and the parent Ab4p viruses without any significant difference (P > 0.05). The growth of the mutant viruses was almost similar across the cell types, but viruses growth was more efficient in FHK cells as judged by the number of the obtained virus particles. The plaque size of Ab4pΔORF68 was significantly (40%) smaller than those of Ab4p (P < 0.01), Ab4pΔORF68R, and Ab4pΔORF68R non-sense viruses which confirmed the importance of ORF68 protein in the cell-to-cell transmission of EHV-1. Subcellular localization of the green fluorescent protein (GFP) ORF68 gene fusion product showed late expression with intranuclear localization of the transfected cells while immunofluorescent antibody technique (IFAT) localized it at the nucleus and nuclear membranes of the infected cells. Hence, it could be concluded that ORF68 protein may not be essential for EHV-1 Ab4p growth but plays a crucial role in virus penetration and transmission at the cellular level. Therefore, the generated EHV-1 ORF68 negative mutant could be a prospective candidate for the development of a vaccine marker.

Clinical, pathological, and molecular data concerning Coenurus cerebralis in Sheep in Egypt

This article contains information related to a recent study “Prevalence and Identity of Taenia multiceps cysts “Coenurus cerebralis” in Sheep in Egypt” (Amer et al., 2017) [1]. Specifically, affected sheep showed neurological disorders manifested as depression, head shaking and circling, altered head position, incoordination and paralysis in some cases. Brain-derived cysts were molecularly identified by PCR-sequence analysis at mitochondrial 12S rRNA gene marker. Cyst-induced pathological changes included degenerative changes and demyelination in brain tissue, infiltration of lymphocytes and histiocytes. Cystic fluids were biochemically analyzed for protein, lipids and electrolytes. The data of this study provides more understanding on phylogeny, epidemiology and pathology of coenurosis in sheep.

Prevalence of yeasts in diarrheic calves with molecular Identification of C. albicans

DOI:10.5176/2382-5685_VETSCI14.29