An Improved PCR-RFLP Assay for Detection and Genotyping of Asymptomatic Giardia lamblia Infection in a Resource-Poor Setting

Laboratory workers, in resource-poor countries, still consider PCR detection of Giardia lamblia more costly and more time-consuming than the classical parasitological techniques. Based on 2 published primers, an in-house one-round touchdown PCR-RFLP assay was developed. The assay was validated with an internal amplification control included in reactions. Performance of the assay was assessed with DNA samples of various purities, 91 control fecal samples with various parasite load, and 472 samples of unknown results. Two cysts per reaction were enough for PCR detection by the assay with exhibited specificity (Sp) and sensitivity (Se) of 100% and 93%, respectively. Taking a published small subunit rRNA reference PCR test results (6%; 29/472) as a nominated gold standard, G. lamblia was identified in 5.9% (28/472), 5.2%, (25/472), and 3.6% (17/472) by PCR assay, RIDA^® Quick Giardia antigen detection test (R-Biopharm, Darmstadt, Germany), and iodine-stained smear microscopy, respectively. The percent agreements (kappa values) of 99.7% (0.745), 98.9% (0.900), and 97.7% (0.981) were exhibited between the assay results and that of the reference PCR, immunoassay, and microscopy, respectively. Restriction digestion of the 28 Giardia-positive samples revealed genotype A pattern in 12 and genotype B profile in 16 samples. The PCR assay with the described format and exhibited performance has a great potential to be adopted in basic clinical laboratories as a detection tool for G. lamblia especially in asymptomatic infections. This potential is increased more in particular situations where identification of the parasite genotype represents a major requirement as in epidemiological studies and infection outbreaks.     

Bovine serum albumin functionalized blue emitting Ti3C2 MXene quantum dots as a sensitive fluorescence probe for Fe3+ ion detection and its toxicity analysis

In the present work, an improved class of protein functionalized fluorescent 2D Ti₃C₂ MXene quantum dots (MXene QDs) was prepared using a hydrothermal method. Exfoliated 2D Ti₃C₂ sheets were used as the starting precursor and transport protein bovine serum albumin (BSA) was used to functionalize the MXene QDs. BSA-functionalized MXene QDs exhibited excellent photophysical property and stability at various physiological parameters. High-resolution transmission electron microscopy analysis showed that the BSA@MXene QDs were quasispherical in shape with a size of ~2 nm. The fluorescence intensity of BSA@MXene QDs was selectively quenched in the presence of Fe³⁺ ions. The mechanism of fluorescence quenching was further substantiated using time-resolved fluorescence and Stern–Volmer analysis. The sensing assay showed a linear response within the concentration range 0–150 μM of Fe³⁺ ions with excellent limit of detection. BSA@MXene QDs probe showed good selectivity toward ferric ions even in the presence of other potential interferences. The practical applicability of BSA@MXene QDs was further tested in real samples for Fe³⁺ ion quantification and the sensor had good recovery rates. The cytotoxicity studies of the BSA@MXene QDs toward the human glioblastoma cells revealed that BSA@MXene QDs are biocompatible at lower doses and showed significant cytotoxicity at higher dosages.     

Eucharitidae (Hymenoptera,Chalcidoidea), a family new to the fauna of Saudi Arabia,with the description of the previously unknown male of Eucharis (Psilogastrellus) affinis Bou?ek

The family Eucharitidae (Hymenoptera: Chalcidoidea) is recorded for the first time for the fauna of Saudi Arabia based on Hydrorhoa caffra (Westwood) and Eucharis (Psilogastrellus) affinis Bouček. The record of Hydrorhoa caffra suggests that Al-Baha and Asir provinces should be considered as part of the Afrotropical rather than the Palaearctic region. The previously unknown male of Eucharis affinis Bouček is described and figured. Macrophotographs of the species are provided.     

Sediment Heavy Metal Contents,Ostracod Toxicity and Risk Assessment in Tropical Freshwater Lakes,Tamil Nadu,India

ABSTRACT

Coimbatore is one of the industrial cities in Tamil Nadu, India, which has been experiencing rapid urbanization and population growth. Coimbatore is also known for its unique freshwater lakes environment and serves as a rich ecosystem. However, the assessment of heavy metal levels in aquatic environments is limited. This study was aimed to investigate physicochemical parameters, heavy metals level and sources, and ecotoxicity in sediments collected from five different lakes in Coimbatore. The concentrations of heavy metals (Cr, Cu, Zn, As, Cd, and Pb) in sediments were determined by Inductive Coupled Plasma-Mass Spectroscopy (ICP-MS). Hg level was measured using Advanced Mercury Analyzer (AMA). The determined heavy metal concentrations in sediments varied significantly according to the lake location and consistent with local human linked anthropogenic activities. The metal concentrations in urban lakes were exceeding both the Sediment Quality Guidelines (SQGs) and the probable effect levels” (PELs) mostly; e.g., in sediments from the Lake Ukkadam, the values of 5.08 and 203.32 mg kg-1 dry weight were observed for Hg and Cu, respectively. The ecotoxicity test with ostracods exposed to the sediment samples revealed that mortality ranged between 6 and 23% for countryside lakes and 28 and 88% for the lakes within urban Zone. We used Spearman rank-order correlation and Principal components analysis (PCA) to assess the sources of pollutants and if they related to anthropogenic pressure and eutrophication of lakes. The main sources of heavy metals from studied lakes differed significantly. Urban and industrial effluents were dominant sources in urban lakes. Agricultural runoff, domestic wastes, and natural weathering were responsible for the metal sources in country lakes. This study provides baseline information on the heavy metal pollution status of sediments in the freshwater lakes in Coimbatore, which will be useful for pollution control measures to prevent possible metal sources on these lakes and impose appropriate management practices and continuous monitoring by relevant authorities.     

Complete chloroplast genome sequences of Dipterygium glaucum and Cleome chrysantha and other Cleomaceae Species,comparative analysis and phylogenetic relationships

This current study presents, for the first time, the complete chloroplast genome of two Cleomaceae species: Dipterygium glaucum and Cleome chrysantha in order to evaluate the evolutionary relationship. The cp genome is 158,576 bp in length with 35.74% GC content in D. glaucum and 158,111 bp with 35.96% GC in C. chrysantha. Inverted repeats IR 26,209 bp, 26,251 bp each, LSC of 87,738 bp, 87,184 bp and SSC of 18,420 bp, 18,425 bp respectively. There are 136 genes in the genome, which includes 80 protein coding genes, 31 tRNA genes and four rRNA genes were observed in both chloroplast genomes. 117 genes are unique while the remaining 19 genes are duplicated in IR regions. The analysis of repeats shows that the cp genome includes all types of repeats with more frequent occurrences of palindromic; Also, this analysis indicates that the total number of simple sequence repeats (SSR) were 323 in D. glaucum, and 313 in C. chrysantha, of which the majority of the SSRs in these plastid genomes were mononucleotide repeats A/T which are located in the intergenic spacer. Moreover, the comparative analysis of the four cp sequences revealed four hotspot genes (atpF, rpoC2, rps19, and ycf1), these variable regions could be used as molecular makers for the species authentication as well as resources for inferring phylogenetic relationships of the species. All the relationships in the phylogenetic tree are with high support, this indicate that the complete chloroplast genome is a useful data for inferring phylogenetic relationship within the Cleomaceae and other families. The simple sequence repeats identified will be useful for identification, genetic diversity, and other evolutionary studies of the species. This study reported the first cp genome of the genus Dipterygium and Cleome. The finding of this study will be beneficial for biological disciplines such as evolutionary and genetic diversity studies of the species within the core Cleomaceae.     

Five new records of bee flies (Bombyliidae,Diptera) from Saudi Arabia with zoogeographical remarks

Five bee-fly species (Bombyliidae, Diptera) have been listed in this paper as new to the Kingdom of Saudi Arabia. Four of the recorded species have been identified to the level of species, namely: Bombomyia discoidea (Fabricius, 1794), Spogostylum candidum (Sack, 1909), Exoprosopa linearis Bezzi, 1924, and Exoprosopa minos (Meigen, 1804), while the fifth one only to genus, Desmatoneura sp. The species have been collected from Al-Baha and Asir Provinces in the south-western part of the Kingdom. One of the four identified species, Exoprosopa linearis, has an Afrotropical affinity, and another two, Spogostylum candidum and Bombomyia discoidea, have considerable Afrotropical distributions, and this result agrees to some extent with studies considering these parts of the Arabian Peninsula, including Al-Baha and Asir Provinces, having Afrotropical influences and may be included in the Afrotropical Region rather than in the Palaearctic Region or the Eremic zone.