Differential detection of echinococcus Spp. copro-DNA by nested-PCR in domestic and wild definitive hosts in moghan plain, Iran

Despite Echinococcus granulosus there are merely two old reports of E. multilocularis infection among Iranian canids of Moghan Plain, the only area known endemic for the species. We detected specific DNA markers in fecal samples by PCR [Copro-PCR] for differential diagnosis of Echinococcus species in living canids. Totally 144 fecal samples from domestic dogs, red foxes and a golden jackal were examined for genus-specific Echinococcus coproantigens using ELISA. Forty two positive or ambiguous samples were further examined for Echinococcus species-specific DNA markers by two different set of nested-PCR. Twenty five out of 144 [17.4%] animals were contaminated with E. granulosus including 14 [23.7%] domestic dogs, 10 [11.9%] red foxes and one [100%] golden jackal. But none of them harboured E. multilocularis species-specific Copro-DNA. The overall prevalence of E. granulosus and E. multilocularis infections in canids of the area was estimated to be 17.4% and 0.0%, respectively. There was a significant relation between the results of Copro-PCR and CA-ELISA. The lack of E. multilocularis infection, compared to previous reports may be due to the differences in used diagnostic methods and/or recently limited territories of wild canids and altered their food resources in this particular area

Comparison of PCR-based diagnosis with centrifuged-based enrichment method for detection of borrelia persica in animal blood samples

The mainstay of diagnosis of relapsing fever [RF] is demonstration of the spirochetes in Giemsa-stained thick blood smears, but during non fever periods the bacteria are very scanty and rarely detected in blood smears by microscopy. This study is aimed to evaluate the sensitivity of different methods developed for detection of low-grade spirochetemia. Animal blood samples with low degrees of spirochetemia were tested with two PCRs and a nested PCR targeting flaB, GlpQ, and rrs genes. Also, a centrifuged-based enrichment method and Giemsa staining were performed on blood samples with various degrees of spirochetemia. The flaB-PCR and nested rrs-PCR turned positive with various degrees of spirochetemia including the blood samples that turned negative with dark-field microscopy. The GlpQ-PCR was positive as far as at least one spirochete was seen in 5-10 microscopic fields. The sensitivity of GlpQ-PCR increased when DNA from Buffy Coat Layer [BCL] was used as template. The centrifuged-based enrichment method turned positive with as low concentration as 50 bacteria/ml blood, while Giemsa thick staining detected bacteria with concentrations >/- 25000 bacteria/ml. Centrifuged-based enrichment method appeared as much as 500-fold more sensitive than thick smears, which makes it even superior to some PCR assays. Due to simplicity and minimal laboratory requirements, this method can be considered a valuable tool for diagnosis of RF in rural health centers

Identification and purification of a specific and immunogenic antigen of the laminated layer of the hydatid cyst and production of an antigen-specific monoclonal antibody

Cystic echinococcosis [CE] is an infection caused by the larval stage of Echinococcus granulosus. This is widely distributed through Iran, where a variety of animals act as intermediate host. The immunogenic antigens [Ag] of different compartments of the hydatid cyst have been already determined. One of these compartments is the laminated layer [LL]. We have extracted a protein with the MW of 24 kDa from a lysate prepared from the LL and produced a monoclonal antibody [mAb] against this protein. Five mAb named P[3]F[6s], P[2]Hp[4s], P[1], A[6s], P[1],C[3s], and P[1],F[7s] have been produced. The isotype analysis showed that P[3]F[6s] is IgG[1], and the rest are IgM. P[3]F[6s] was purified from the ascitic fluid of mice injected with P[3]F[6s] hybridoma intraperitoneally. Western blot and LLISA analysis showed that this mAb could recognize the purified 24 kDa prepared from lysate of LL. Since a 24 kDa protein has been shown to be an immunogenic Ag, this protein can be used as a candidate for the development of diagnostic tests and vaccine strategies. For these aims, P[3]F[6s] can be used for the purification of this 24 kDa protein

Prevalence of zoonotic intestinal helminths of canids in Mogahan plain, Northwestern Iran

The present study was aimed to elucidate the status of intestinal helminth infections in canids of Moghan Plain, northwestern Iran. Eighty-five intestine samples from dead or shot wild canids, 59 fecal samples from sheepdogs and 5 from red foxes were collected from 2006 to 2008 and examined in Parasitology department of Pasteur Institute of Iran. Generally, adult worms, larvae, and eggs of 13 species of various parasitic helminths were recovered. Necropsy examinations showed that 96.47% animals harbored at least one helminth species. The prevalence of different species in necropsy were Mesocestoides sp. 84.7%, Rictolaria spp. 55.3%, Macranthorhynchus hirudinaceus 45.9%, Toxocara canis 43.5%, Toxascaris spp. 35.3%, Joyeuxiella sp. 34.1%; hookworms; 22.4%, Taenia spp. 11.8%, Alaria spp. 2.4% and Dipylidium caninum 1.2%. Besides, eggs belonging to 10 species of parasitic helminths were identified in 46 fecal samples and generally, 30.9% of samples harbored eggs of at least one helminth species. The high prevalence of various helminth infections among canids in Moghan plain and contamination of environment by helminths eggs may increase the risk of infection for native people

Characterization of specific IgE antibody related to antigen 5 of echinococcus granulosus

Anaphylactic reactions, such as urticaria, edema, respiratory symptoms, and anaphylactic shock often complicate the course of Cystic Echinococcosis [CE]. To investigate the role of the IgE immunoreactive antigen 5 [Ag 5] in the sero-positive patients with CE, we determined N-terminal of 57 kDa subunit of Ag5 responsible for IgE and C-terminal of this active antigen related to induction of IgG specifically. Immunoblotting analysis showed that specific IgE to 57-kDa subunit related to inter-chain disulphide band of two 22 kDa and 38-kDa component of Ag5 and conformational epitope on this subunits. In addition, since the 57 kDa component arise from the removal of the C-terminal portion of 22 kDa subunit of Ag5, thus IgE specifically recognized N-terminal of 22 kDa subunit which remain bounds to the other component, whereas IgG reacted with C-terminal of 38 kDa component of Ag5. Recognition of the specific binding site on the 57 kDa subunit of Ag5 could leads to understanding the mechanism regulating IgE/IgG production in some immune circumstances that IgE tends to some dominate, whereas in other IgG predominates

Nasopharyngeal pentastomiasis [Halzoun]: report of 3 cases

A three-member family from Tehran referred with variety of nasopharyngeal symptoms including sneezing, coughing and nasal discharge following consumption of barbecued liver [Kabab]. Thirteen worm-like nymphs of Linguatula serrata were separated from larynx, nose and gum of these patients. The adult parasites live in the nose and paranasal sinuses of dogs and other carnivores. Although this infection is rare, in population who have the habit of consuming undercooked internal organs of herbivorous animals the physicians should consider its probability