Background and Aims: The aim of this study is to clarify nitric oxide (NO)-production by spleen and the importance of spleen in malaria infection in murine model.
Materials and Methods: Thirty outbred NMRI female mice were divided into four groups, Group I: No intervention (Healthy control), Group II: With splenectomy (Healthy test), Group III: No intervention, Inoculation of contaminated blood (Infected control), Group IV: With splenectomy, inoculation of contaminated blood (Infected test). The Parasitemia was counted every other day through Giemsa stain examination of animal blood. The parasitemia and survival rates, hepatosplenomegaly and body weight were recorded. After terminal anesthesia, plasma and liver/spleen suspensions were assessed by the Griess micro assay for measurement of NO-levels.
Results: At the end of the experiment (on day 16), the parasitemia was 26.99±0.46 % among the group of non-splenectomized animals (Group III) compared with 31.25±0.72% among the group of splenectomized animals (Group IV). The average parasitemia among the groups at the end of the experiment was statistically significant (Group III, Group IV: p= 0.0002). Survival rate was statistically significant (p<0.0001). NO concentrations in plasma, liver and spleen were determined. The amount of NO in plasma increased significantly in the infected groups (p=0.0003).
Conclusions: Although, splenectomy decreased immune function against rodent malaria, it did not solely changed the pattern of antimalarial activity via NO-pathway. It is concluded that NO possibly comes from several sources rather than spleen during rodent malaria disease and is released into circulation, which may replace NO shortage by splenic cells to combat malaria parasites.
Background
Dicrocoelium dendriticum is a broadly distributed zoonotic helminth, which is mainly reported from domesticated and wild ruminants. There is little data covering the molecular features of this trematode; therefore, current study aimed to molecularly analyze D. dendriticum in livestock.
Methods
Totally, 23 samples of D. dendriticum were collected from cattle, sheep, and goat from Ilam, Lorestan, and Khuzestan, three west and south-west provinces of Iran from February to August 2018. After genomic DNA extraction, the internal transcribed spacer (ITS) 2 fragment was amplified and sequenced in samples. To investigate genetic variations through the ITS 2 fragment of obtained D. dendriticum, phylogenetic tree and network analysis were employed.
Results
All 23 samples were successfully amplified and sequenced. Phylogenetic tree showed that our samples were clearly grouped in a clade together with reference sequences. There was no grouping based on either geographical regions or hosts. Network analysis confirmed the phylogenetic findings and showed the presence of nine distinct haplotypes, while our samples together most of sequences, which were previously submitted to the GenBank, were grouped in the Hap1.
Conclusions
Our findings indicated that although ITS 2 fragment discriminate D. dendriticum, this fragment is not suitable to study intra-species genetic variations. Therefore, exploring and describing new genetic markers could be more appropriate to provide new data about the genetic distribution of this trematode.
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