Allopatric speciation in the desert: diversification of cichlids at their geographical and ecological range limit in Iran

Collated electrofishing data from wadeable riffles of boreal rivers in Finland revealed a substantial shift in the fish assemblage composition, accompanied by a decline in total fish density and a reduction in species richness from early summer to late autumn. As the major changes in fish assemblages, the density of cyprinids decreased sharply from June towards autumn, whereas salmonids peaked in September. These shifts were considered to originate mainly from spawning migrations, fish movements to winter refuge habitat and other temperature-related responses of rheophilic versus eurytopic fish species. Temporal change in fish assemblages induced a prominent variation in the monthly fish-based index values used for bioassessment. The proportion of sampled sites classified as high or good in ecological status within the Water Framework Directive (WFD) based on fish increased from 25.9% in July to 68.3% in October. These results, combined with the observed timing of young of the year (0+) fish recruitment to electrofishing catch, suggest that sampling of fish in boreal rivers for WFD monitoring should be restricted to a considerably shorter period than the prevailing practice in order to avoid temporal bias.     

Phylogenetic relationships and taxonomy of <Emphasis Type="Italic">Luciobarbus barbulus</Emphasis> (Heckel, 1847) (Teleostei: Cyprinidae)

Luciobarbus barbulus (Heckel, 1847), a cyprinid fish of the Persian Gulf basin is re-described. Its identity is discussed and its phylogenetic relationship among the tribe Barbini is reconstructed. Luciobarbus barbulus can be distinguished from its close species, L. pectoralis by having thicker lower lip, stronger last unbranched dorsal fin ray and with stronger serration, longer barbels and posterior barbel passes the middle of the eye (vs. not reaching the middle of the eye in L. pectoralis), confirming the identity of both, as distinct species. Moreover, phylogenetic analysis based on the mtCOI sequences, placed the sequenced L. barbulus in a distinct clade corresponding to the tribe Barbini which includes Aulopyge, Barbus and Luciobarbus (three genera from Eurasia and North of Africa), sister to L. pectoralis from Orontes River with genetic distance of 1.7% applying the Kimura 2 Parameter (K2P) model.     

Detection of adulteration in Iranian saffron samples by <Superscript>1</Superscript>H NMR spectroscopy and multivariate data analysis techniques

Introduction

The high market value of saffron (Crocus sativus L.) has made it an attractive candidate for adulteration. Safflower (Carthamus tinctorius L.) and tartrazine are among the most common herbal and synthetic foreign materials that may be added to pure saffron for the purpose of adulteration. In spite of encouraging advances achieved in the identification of adulteration in saffron samples, the lack of a simple method with sufficient power for discrimination of pure high grade saffron from meticulously adulterated saffron samples persuaded us to perform this study.

Objectives

In this work, we show that ¹H NMR spectroscopy together with chemometric multivariate data analysis methods can be used for the detection of adulteration in saffron.

Methods

Authentic Iranian saffron samples (n?=?20) and adulterated samples that were prepared by adding either different quantities of natural plant materials such as safflower, or synthetic dyes such as tartrazine or naphthol yellow to pure saffron (n?=?22) composed the training set. This training set was used to build multivariate Principal Component Analysis (PCA) and Partial Least Squares Discriminant Analysis (PLS-DA) models. The predictive power of the PLS-DA model was validated by testing the model against an external dataset (n?=?13).

Results

PCA and PLS-DA models could both discriminate between the authentic and adulterated samples, and the external validation showed 100% sensitivity and specificity for predicting the authenticity of suspicious samples. Peaks specific to authentic and adulterated samples were also characterized. Proximity of samples with unknown adulteration status to the samples adulterated with known compounds in the PCA provided insight regarding the identity of the adulterant in the suspicious samples. Furthermore, the authentic samples could be distinguished based on their cultivation site.

Conclusion

The present study demonstrates that the application of ¹H NMR spectroscopy coupled with multivariate data analysis is a suitable approach for detection of adulteration in saffron specimens. Outstanding sensitivity and specificity of the PLS-DA model in discriminating the authentic from adulterated samples in external validation confirmed the high predictive power of the model. The advantage of the present method is its power for detecting a wide spectrum of adulterants, ranging from synthetic dyes to herbal materials, in a single assay.

     

Variation of DAT1 VNTR alleles and genotypes among old ethnic groups in Mesopotamia to the Oxus region

Variation of a VNTR in the DAT1 gene in seven ethnic groups of the Middle East was used to infer the history and affinities of these groups. The populations consisted of Assyrian, Jewish, Zoroastrian, Armenian, Turkmen, and Arab peoples of Iran, Iraq, and Kuwait. Three hundred forty subjects from these seven ethnic groups were screened for DAT1. DAT1 VNTR genotyping showed 3, 6, 7, 8, 9, 10, 11, and 12 alleles in the samples. Analysis of these data revealed differentiation and relationship among the populations. In this region, which covers an area of 2-2.5 million km2, the influence of geography and especially of linguistic characteristics has had potentially major effects on differentiation. Religion also has played a major role in imposing restrictions on some ethnic groups, who as a consequence have maintained their community. Overall, these ethnic groups showed greater heterogeneity compared to other populations.     

Reproductive biology of the endemic Iranian cichlid, <Emphasis Type="Italic">Iranocichla hormuzensis</Emphasis> Coad, 1982 from Mehran River,southern Iran

We studied the reproductive biology of the endemic Iranian cichlid Iranocichla hormuzensis, a maternal mouthbrooder, in the Mehran River. Overall sex ratio was 1.44 M: 1F indicating a significant male bias. Monthly sex ratios did not differ from unity except in May and June when the sex ratios were strongly biased in favor of males. Four different reproductive indices and the high frequency of large oocytes, all suggest that the Iranian cichlid spawns at the end of winter and the beginning of spring. Mircoscopic gonadal maturation stages for both males and females were correlated to the gonadosomatic indices. The maximum number of larvae in the mouth of a female was 153.     

The relationship between established coronary risk factors and serum copper and zinc concentrations in a large Persian Cohort

IntroductionThe relationship between demographic and biochemical characteristics, including several established coronary risk factors, and serum copper and zinc was assessed in a large Iranian population sample.Materials and methodsA group of 2233 individuals, 15–65 years of age [1106 (49.5%) males and 1127 (50.5%) females] was recruited from residents of the Greater Khorasan province in northeast of Iran. Demographic data were collected using questionnaires. Coronary risk factors were determined using standard protocols, and trace elements were measured in serum using atomic absorption spectroscopy.ResultsDegree of glucose tolerance and smoking habit were not associated with serum zinc and copper levels. Serum copper levels were significantly higher in obese and hypertensive than in normal subjects (p<0.001). In the whole group and for the female subgroup, serum zinc (p<0.01) and copper (p<0.001) were both significantly lower in individuals with normal versus high levels of low-density lipoprotein cholesterol.A strong positive correlation was found between serum copper and body mass index (BMI) (r=0.85, p<0.001). Weaker positive associations were found between serum copper and calculated 10 years’ coronary risk (r=0.11, p<0.001). Serum zinc/copper ratio was strongly inversely associated with calculated 10 years’ coronary risk (r=?0.10, p<0.001). The partial Eta squared (PES) values for factors determining serum zinc were hypertension (0.007, p=0.01) and BMI (0.004, p=0.01); and for serum copper, they were gender (0.02, p=0.001), hypertension (0.004, p=0.009), and 10 years’ coronary risk for men (0.003, p=0.03) and women (0.002, p=0.07).ConclusionSignificant associations between serum trace element concentrations and several coronary risk factors, including calculated 10 years’ coronary risk scores, were found.     

PCR-based detection and eradication of mycoplasmal infections from various mammalian cell lines: a local experience

A total of 200 cell lines including different human, monkey, mice, hamster and rat cell types were examined for mycoplasma infection status. PCR assay using generic-specific universal primers showed that 40 (20%) of the cell lines are contaminated with mycoplasma. Employment of species-specific primers within these infected cell lines revealed infection with M. hyorhinis (42.5%), M. fermentas (37.5%), M. arginini (37.5%), M. orale (12.5%) and A. laidlawii (7.5%). A number of the cultures were coinfected with 2 or 3 different species. Contaminated samples were treated with BM-Cyclin, Ciprofloxacin and mycoplasma removal agent (MRA). Mycoplasma eradication was subsequently checked by PCR following 2 weeks continuous culture of treated cells in antibiotic free culture medium. Mycoplasmal infections were eradicated in 100, 70 and 42% of infected cell lines when the samples were treated with BM-Cyclin, MRA and Ciprofloxacin, respectively. However, 12% (BM-Cyclin), 62.5% (MRA) and 82.5% (Ciprofloxacin) of mycoplasma regrowth was observed 4 months after the treatment. Notably, the risk of spontaneous culture death was 17.5, 12.5 and 0% for BM-Cyclin, MRA and Ciprofloxacin, respectively.     

Killing of Trypanosomatid Parasites by a Modified Bovine Host Defense Peptide,BMAP-18

Background

Tropical diseases caused by parasites continue to cause socioeconomic devastation that reverberates worldwide. There is a growing need for new control measures for many of these diseases due to increasing drug resistance exhibited by the parasites and problems with drug toxicity. One new approach is to apply host defense peptides (HDP; formerly called antimicrobial peptides) to disease control, either to treat infected hosts, or to prevent disease transmission by interfering with parasites in their insect vectors. A potent anti-parasite effector is bovine myeloid antimicrobial peptide-27 (BMAP-27), a member of the cathelicidin family. Although BMAP-27 is a potent inhibitor of microbial growth, at higher concentrations it also exhibits cytotoxicity to mammalian cells. We tested the anti-parasite activity of BMAP-18, a truncated peptide that lacks the hydrophobic C-terminal sequence of the BMAP-27 parent molecule, an alteration that confers reduced toxicity to mammalian cells.

Methodology/Principal Findings

BMAP-18 showed strong growth inhibitory activity against several species and life cycle stages of African trypanosomes, fish trypanosomes and Leishmania parasites in vitro. When compared to native BMAP-27, the truncated BMAP-18 peptide showed reduced cytotoxicity on a wide variety of mammalian and insect cells and on Sodalis glossindius, a bacterial symbiont of the tsetse vector. The fluorescent stain rhodamine 123 was used in immunofluorescence microscopy and flow cytometry experiments to show that BMAP-18 at low concentrations rapidly disrupted mitochondrial potential without obvious alteration of parasite plasma membranes, thus inducing death by apoptosis. Scanning electron microscopy revealed that higher concentrations of BMAP-18 induced membrane lesions in the parasites as early as 15 minutes after exposure, thus killing them by necrosis. In addition to direct killing of parasites, BMAP-18 was shown to inhibit LPS-induced secretion of tumour necrosis factor alpha (TNF-α), a cytokine that is associated with inflammation and cachexia (wasting) in sleeping sickness patients. As a prelude to in vivo applications, high affinity antibodies to BMAP-18 were produced in rabbits and used in immuno-mass spectrometry assays to detect the intact peptide in human blood and plasma.

Conclusions/Significance

BMAP-18, a truncated form of the potent antimicrobial BMAP-27, showed low toxicity to mammalian cells, insect cells and the tsetse bacterial symbiont Sodalis glossinidius while retaining an ability to kill a variety of species and life cycle stages of pathogenic kinetoplastid parasites in vitro. BMAP-18 also inhibited secretion of TNF-α, an inflammatory cytokine that plays a role in the cachexia associated with African sleeping sickness. These findings support the idea that BMAP-18 should be explored as a candidate for therapy of economically important trypanosome-infected hosts, such as cattle, fish and humans, and for paratransgenic expression in Sodalis glossinidius, a bacterial symbiont in the tsetse vector, as a strategy for interference with trypanosome transmission.     

Portable,Battery-Operated,Low-Cost,Bright Field and Fluorescence Microscope

This study describes the design and evaluation of a portable bright-field and fluorescence microscope that can be manufactured for $240 USD. The microscope uses a battery-operated LED-based flashlight as the light source and achieves a resolution of 0.8 µm at 1000× magnification in fluorescence mode. We tested the diagnostic capability of this new instrument to identify infections caused by the human pathogen, Mycobacterium tuberculosis. Sixty-four direct, decontaminated, and serially diluted smears were prepared from sputa obtained from 19 patients suspected to have M. tuberculosis infection. Slides were stained with auramine orange and evaluated as being positive or negative for M. tuberculosis with both the new portable fluorescence microscope and a laboratory grade fluorescence microscope. Concordant results were obtained in 98.4% of cases. This highly portable, low cost, fluorescence microscope may be a useful diagnostic tool to expand the availability of M. tuberculosis testing at the point-of-care in low resource settings.     

Comparative analysis of essential oil compositions in seven populations of Bakhtiarian savory in natural and field conditions

Bakhtiarian savory (Satureja bachtiarica Bunge) is an endemic herb growing wild in Iran with interesting pharmacological and biological properties. In this research, the variability of essential oil content and phytochemical compositions among seven populations of this species was studied in both natural and field conditions. There was a wide interpopulation variability in phytochemical variation of studied populations under both natural and field conditions, indicating the existence of different chemotypes. Six chemical compositions showed high value in studied plants so that carvacrol and thymol were known as major compounds and confirmed by principal component analysis. Ilam population had the highest thymol in both field (77.10 %) and natural (42.32 %) conditions and was known as a thymol chemotype. Beside, Yazd and Shahrekord populations, respectively, showed the highest carvacrol percentages in both field (84.83 and 63.81 %) and natural (72.50 and 58.05 %) conditions and were known as carvacrol chemotypes. In other populations, carvacrol and thymol were the main components and showed partly equal percentage and these populations were known as carvacrol/thymol chemotypes. According to Pearson correlation analysis, carvacrol was negatively correlated with p-cymene, γ-terpinene and thymol (r = ?0.61, ?0.70 and ?0.98, respectively), indicating very high reverse correlation between carvacrol and thymol. In conclusion, the high value of the essential oil compositions in studied S. bachtiarica populations could provide useful information for conservation and selection of cross parents in breeding programs to develop improved cultivars.