Genotypic and phenotypic analysis of Fasciola isolates

To identify the fasciolid species by morphometric and molecular methods in Zanjan, northwest of Iran. Adult Fasciola worms [n=584] were obtained from cattle and sheep in Zanjan slaughterhouse in 2007. Living flukes were washed, then worms' images were taken by 3CCD camera and finally apical zone of each worm was obtained. Morphometric values such as body length, body width, body area, body perimeter and the distance between ventral sucker and posterior end of body were obtained using AutoCAD image analysis software. Total gDNA was extracted from individual flukes by modified phenol-chloroform method. PCR amplification of ITS2 fragment was performed the isolated DNA samples and the amplicons were consequently subjected to RFLP assay and nucleotide sequencing to distinguish between fasciolid species. Mean of morphometric values in flukes from sheep was greater than those of cattle. Accordingly, the identified species included 31% F. hepatica-like, 7% F. gigantica-like and 62% intermediate forms. However, ITS2 fragment of 535 amplified specimens, showed no variation at the species-specific nucleotide sites 230, 340 and 341. The amplified fragment composed of partial 5.8S sequence [62bp], the complete ITS2 sequence [361bp] and partial 28S sequence [34bp]. The nucleotide contents of ITS2 region were 69 A, 116 T, 81 C and 95 G and the average G+C content was approximately 49%. Comparing of ITS2 sequences with the BLAST GenBank database, also confirmed that all specimens were F. hepatica. A simple and rapid PCR-RFLP assay can be used for distinguishing between these species

Immunoregulatory cytokine [TGF-beta and IL-10] responses in mice inoculated with protoscoleces and major hydatid fluid antigens of cystic echinococcosis

Our objectives were to investigate whether immunomodulatory cytokines, TGF-beta and IL-10, are stimulated in response to cystic echinococcosis [CE] components in mice model, and whether major hydatid fluid antigens or live protoscoleces could equally contribute to such cytokines. Protoscoleces were obtained by aseptic puncture of fertile sheep hydatid cysts. Hydatid fluid antigens [HFAgs] and Antigen B [AgB] were prepared by partial purification and electroelution, respectively. Of the 25 Balb/c mice assigned in four groups, the first group was inoculated ip with 2000 live protoscoleces; the second and the third groups were injected ip with 50 micro g HFAgs and 50 micro g AgB in 200 micro l of PBS, respectively. Control group was only injected with PBS. The sera concentration of TGF-beta and IL-10 were determined by ELISA. Data were analyzed using One-Way ANOVA and Tukey-HSD tests to compare differences between means. The mean concentration of TGF-beta in those groups injected with protoscoleces, HFAgs and AgB were significantly higher than control group. However, in the case of IL-10 such differences were only detected in mice that were inoculated with protoscoleces [356 +/- 11 pg/ml] compared to control [207 +/- 9 pg/ml], HFAgs and AgB groups. TGF-beta and IL-10, two important immunomudulatory cytokines are induced by different molecules or components of CE, so that AgB could induce TGF-B and components of protoscolex, other than AgB and Ag5, could induce IL-10

fauna and frequency of different mosquito species [diptera: culicidae] in Zanjan Province, 2002-2003

Mosquitoes are the most important blood feeding insects with high health and medical significance. Zanjan province forms one of the most important permanent mosquito habitats. With regard to different aspects during past three decades, antiquity of past surveys, necessity of control programs against mosquito-borne diseases, this study was conducted to determine the fauna of mosquito species in Zanjan during 2002-2003. In this descriptive study the adult mosquitoes were collected from 5 fixed indoor and outdoor areas near larval habitats of Zanjan, Mahneshan, Tarom, Abhar and Khodabandeh districts. Sampling was carried out ten times at intervals of 15 days during September through October [2002] and June through August [2003] by pyrethrum space spray [PSS] and light traps [LT], while larvae were collected by hand catch. Collected samples were examined under microscope and identified according to standard keys. In this study 16743 mosquitoes were collected using PSS method. Over 74% of the collected mosquitoes [12549] were Anopheles maculipennis complex and 21% [3520] Culex theileri; Cx. pipiens, Culiseta longiareolata. An. superpictus and Cs. subochrea were the other mosquito species and consisted of 4.2% [674] of the samples. Around 3723 larvae [3-4th instars] were also collected from constant larval habitats. Cx. theileri larvae included 68.25% [2541] while approximately 16% of the larvae [607] belonged to An. maculipennis complex, An. superpictus 8.4% [313], Cs. longiareolata 3.2% [132], Cx. laticinctus 2.8% [90] and Cx. hortensis 0.8% [29]. An. hyrcanus and Cs. subochrea were also found scarcely which numbered 9 and 2 respectively. Like PSS method, 47273 adult mosquitoes were collected by light trap from July to September, out of which 87.92% were Cx. theileri and 11.12 An. Maculipenins. Ecologic and molecular studies on various populations of Anopheles maculipennis, An. Superpictus, and Culex theiler are recommended. The Species An. Hyrcanus, Cx laticinctus Cx. Hortensis, Cs. longiareolata and Cs. Subochrea species should be taken into consideration in differential diagnosis of the provinces' species

Estimation and comparison of anopheles maculipennis s.l. [Diptera: Culicidae]survival rates with light-trap and indoor resting data

The main vector of malaria in Europe and Palearctic region is Anopheles maculipennis complex. In order to determine the survival rate of An. maculipennis s.l. this study was carried out in Garaboteh [Zanjan province, Iran] in July 2003. In this study anophelines were sampled for 31 consecutive days from five fixed animal shelters and five light traps. Out of 24481 collected An. maculipennis s.L, 19703 [80.48%] were female. Relative density of female anopheline with 95% CI was 74.25-78.48 and 49.16-52.36 in light traps [LT] and pyrethrum space spray catch [PSC], respectively. A significant difference was in mean parous rate in LT [0.46] and PSC [0.50] samples [P< 0.01]. Daily survival rate of anopheline mosquitoes in study area was 0.82-0.86. About 1.74% of female mosquitoes can transmit malaria after 10 d. In 4 d gonotrophic cycle, there was maximum correlation [>0.92] between parous and total females. Also there was significant correlation between nulliparous females and males in LT [r=0.96] and PSC [r=0.87] samples and with increasing anopheline abundance ratio of male/nulliparous significantly decreased [r=-0.35, P<0.05]. Due to simplicity and feasibility of using graphical and parametric method, it can be placed in entomological surveys for monitoring parity, age structure and survival rate of vector anopheline in the world

Anopheline sporozoite infection rate in ghasseghand, baluchis, Iran [1995]

In the study carried out in the ghassreghand district [Baluchistan Iran] from March through November, 1995, a 2 site Sandwich Enzyme - Linked Immunosorbent Assays [ELISA5] were used to identify Plasinodjum falciparum and P.vivax sporozoite infection in 6 Anopheles species which collected monthly indoor resting habitats of 13 villages by "Pyrethrum spray catch" procedure. EIISA tests on 9016 mosquitoes detected 1.5% [53 of 3510] of Anopheles cuIjcif…cies, 0.27% [14 of 5090] of An. stephensi, 3.5% [8 of 226] of An. pulcherrimus and 0.67% [1 of 50] of An. dthali were positive for P. falciparum and P. vivax sporozojte. Positive ELISA results revealed, seasonal malaria transmission in this part of Baluchjstan is from April through early November, and anopheline sporozoite rate is negatively by correlated with anopheline indoor resting density. In addition, An. culicifacies is considered as the main, and the rest of Anopheles species are secondary vectors of malaria in this highly endemic district