In vitro and in vivo anthelmintic efficacy of condensed tannins extracted from the seeds of alfalfa (Medicago sativa L.) against Haemonchus contortus infection

This study was designed to examine in vitro and in vivo anthelmintic efficacy of condensed tannins (CT) extracted from seeds of Medicago sativa on Haemonchus contortus in sheep. CT's in vitro anthelmintic effect was assessed at a 300 μg/ml concentration compared with albendazole (reference drug) at 10 μg/ml. The results showed that CT had a nematocidal effect on H. contortus, and the cuticle of the adult worm appeared to be its initial target. For the in vivo experiment, nine 3-month-old helminths-free lambs were distributed into three groups. Group 1 (n=3) was challenged only as infected untreated controls; Group 2 (n=3) was treated with condensed tannin, and Group 3 (n=3) was treated with albendazole. Fecal and blood samples were collected every 3 days until the end of the experiment; for fecal egg count (FEC) and anti- H. contortus IgG titers determination, respectively. The lambs treated with the CT in G2 exhibited a pronounced decrease of mean FEC with great FECR% detected from the first-week post-treatment (PT) until the end of the experiment compared with G1 animals. The antibody levels gradually increased in G2 following the 2nd dose of CT treatment compared to other groups. A brilliant consistent relation between the elevation of IgG response and reduction of FEC was observed following the second booster dosing of the CT in G2. In conclusion, the CT evoked strongly in vitro and in vivo anthelmintic activity against H. contortus and could be used as a natural alternative treatment of high potency against haemonchosis in sheep

Th1/Th2 balance and humoral immune response to potential antigens as early diagnostic method of equine Strongylus nematode infection

Aim: The aim of this study was to investigate the early diagnosis of strongyle infection based on early changes in Th1 and Th2 cytokines beside the diagnostic accuracy values and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting profiles using prepared strongyles antigens. Materials and Methods: A total of 73 donkeys had a mean age of 4-32 years old were parasitologically examined for strongyle infection. The early changes in Th1 and Th2 cytokines were determined, and the diagnostic accuracy values and SDS-PAGE and western blotting profiles were performed using prepared strongyles antigens; crude somatic Strongylus vulgaris (CSS), excretory-secretory S. vulgaris (ESS), crude somatic Cyathostomins (CSC), and excretory-secretory Cyathostomins (ESC). Results: The results revealed highest 437.04% and lowest 37.81% immunoglobulin G (IgG) in high and low egg shedder groups when using ESC and CSS antigens, respectively. Antibodies index for ESS and CSC were significantly higher in moderate egg shedder group while that for ESS and CSC, ESC was significantly higher in high egg shedder group. Tumor necrosis factor alpha (TNF-α)/interleukin-4 (IL-4) balance in S. vulgaris infected donkeys was approximately equal in apparently healthy, low and high egg shedder groups while TNF-α < IL-4 in moderate egg shedder. In Cyathostomins infected animals, TNF-α/IL-4 balance was approximately equal in apparently healthy group while it was low in moderate and high egg shedder groups. The diagnostic accuracy showed that the higher specificity (46.6%) and prevalence (95.40%) were recorded by CSS and ESC antigens, respectively. However, SDS-PAGE and western blotting profiling proved that the band at molecular weight 25 kDa is exhibited by CSS antigen. Conclusion: Combination of detecting level of TNF-α/IL-4 balance, CSS antigen and IgG concentration is good tool for appropriate diagnosis of such infection. More advancement research must be done concerning Th1/Th2 balance and cross-reactivity of S. vulgaris and Cyathostomins spp. at the base of serological and molecular investigation

Emergent and Neglected Equine Filariosis in Egypt: Species Diversity and Host Immune Response

Equine filariosis (EF) is a neglected vector-borne disease caused by nematode species belonging to the Onchocercidae and Setariidae families. Aside from their zoonotic potential, some species are responsible for serious health problems in equids worldwide, leading to significant economic difficulties. Here, we molecularly investigated equine blood samples (320 horses and 109 donkeys from Egypt) and four adult worms isolated from the peritoneal cavity of 5 out of the 94 slaughtered donkeys. In addition, quantitative enzyme-linked immunoassays (ELISAs) targeting circulating cytokines were used to identify whether the immunological profile of the infected animals is a Th1 (i.e., INF-gamma as indicator) or Th2 (i.e., IL-5 and IL-10 as indicators) response type. Overall, 13.8% and 0.3% of the donkeys and horses, respectively, were scored as positive for filaroid DNA. The 18S phylogeny revealed the occurrence of three different filaroid species, identified here as Mansonella (Tetrapetalonema) sp., Setaria digitata and Dirofilaria repens. Th1 (INF-gamma and IL-5) and Th2 (IL-10) immune response types were identified in equines infected with S. digitata and Mansonella (T.) sp., respectively. These results provide new data on the species diversity of EF in Egypt and extend knowledge of the downregulation of the protective immune response by the potentially zoonotic Mansonella (T) sp. There is an urgent need to implement control measures to preserve equine health and limit the propagation of these vector-borne filaroids in Egypt