Introduction Eimeria spp. are intracellular protozoan parasites of the phylum Apicomplexa causing economic losses to various wild and domestic animals. An eimerian species infecting Columba livia ...domestica was identified in this study. Methods A total of 15 faecal samples were examined by floatation technique, a prevalence rate of 60% was reported. Eimerian oocysts were sporulated in 2.5% potassium dichromate solution then identified using morphological and molecular (DNA amplification of the 18S rRNA and ITS-1 genes) diagnostic techniques. Results Sporulated oocysts were identified as Eimeria labbeana -like, after morphometry with typical bi-layered wall with spherical to subspherical oocysts morphology. A polar granule is present, but no micropyle or oocyst residuum. Sporocysts are elongated ovoidal with stieda body. Sporocyst residuum with many granules and sporozoites with refractile bodies and nucleus. Both 18S rRNA and ITS-1 sequences have been deposited in GenBank database. DNA sequences from the partial 18S rRNA generated from the oocysts were found to be related to eimerian and isosporan parasites found in domestic pigeons. For the first time, ITS-1 sequences for E. labbeana -like were provided. Conclusion The necessity of using molecular techniques to describe pigeon intestinal coccidian parasites in conjunction with traditional morphology-based tools was emphasized in this work in order to understand the biology of such parasites.
•Novel techniques are encouraged to identify nontuberculosis mycobacteria (NTM).•An algorithm incorporating direct molecular identification of NTM is suggested.•Rapid NTM identification can assist in ...decision-making and prompt treatment initiation.
Nontuberculous mycobacteria (NTM) are a group of acid-fast mycobacteria other than Mycobacterium tuberculosis complex (MTBC) that cause pulmonary disease that is similar to the disease caused by MTBC. International guidelines for the diagnosis of pulmonary NTM disease are rigid and have remained unchanged for nearly 2 decades. In this opinion piece, we provide a new perspective on the traditional criteria by suggesting a diagnostic algorithm that incorporates direct molecular identification of NTM performed on raw sputum specimens (using Sanger or targeted deep sequencing approaches, among others) paired with traditional culture methods. Our approach ensures a more rapid diagnosis of pulmonary NTM disease, thus, facilitating timeous clinical diagnosis, and prompt treatment initiation, where indicated, and leverages recent advances in novel molecular techniques into routine NTM identification practice.
The development of polymerase chain reaction (PCR) markers to identify theY chromosome of Ceratitis capitata Wiedemann has permitted the detection of sperm transferred to females during mating. ...However, a molecular technique to quantify the sperm transferred has not yet become available. The current method to quantify the amount of sperm has been the direct counting of sperm heads. Thus, the purpose of this research was to develop and validate an accurate molecular method of diagnosis based on the application of an absolute quantitative real-time PCR, which allows the assessment of the quantity of sperm stored in the spermathecae. For this, Y-specific sequences were used to re-design and test distinct sperm markers. From the amplification product of samples detected as strong positives in conventional PCR, a cloning process of the target sequence was carried out to build the required standard curve. A series of known dilutions of this standard material was prepared for the absolute quantification process. A Roche Lightcycler 480 Real-Time PCR System and SYBRGreen fluorescent dye were used to quantify the sperm contained in the spermathecae of 4-d-old mated females and virgins. Wild-type and Vienna-8 strain sterile males were used to quantify the sperm transferred at four mating durations (10, 30, 60, and 90 min) under laboratory conditions. To validate the reported quantitative method, our results were compared by counting sperm heads under a fluorescent microscope using the same experimental design. In addition, DNA samples were also evaluated and compared by conventional PCR.
This study aims to investigate some chemical, physical, textural, and aroma compounds properties of the ice-cream with berry fruits produced using classical and molecular ice-cream production ...techniques. No significant difference was determined between the pH, dry solid, fat, protein, ash amounts, and overrun of the samples. However, molecular ice-creams, in particular, have a softer structure since they contain tiny ice crystals, and the first dripping and full melting time was realized in less time. The effect of added fruit variety on the viscosity value of the samples was found to be significant. It was observed that the volatile compound profiles of the ice-creams were affected differently by ice-cream production techniques. The octanoic, hexanoic, and dodecanoic acid were high-value prominent compounds in all the ice creams. Furthermore, the 14 volatile compounds that are effective in the difference between the ice-creams and their production techniques were determined, which is more important in the ice cream aroma. This is the first study on the change of the volatile compounds, hence the aroma compound profile, in molecular ice-cream production technique. And, it was presented as an innovative approach. Therefore, it is thought to be shed new light on future studies.
Background: Demodicosis, also named as demodectic mange, red mange or follicular mange (Shrestha et al., 2015). In dogs, Demodex canis is acquired from the dam during the first few hours of life, ...probably during suckling (Greve and Gaafar, 1966). Demodicosis can be defined on the basis of two forms localized and generalized (Shipstone, 2000) with juvenile or adult onset. The ideal confirmation of diagnosis of demodicosis were established by the laboratory analysis of the cutaneous skin scrapings. Various drugs have been used for treating canine demodicosis. Till date no research work has been done on herbal nano medicine against demodicosis in dogs especially in Mizoram. Keeping these points in view the present study was made to formulate the herbal nanomedicine against demodicosis in dogs.Methods: Present investigation was conducted for curing canine demodicosis with the help of herbal essential oils along with its ameloriation with silver Nanoparticles. A total of 1200 dogs were screened for canine demodicosis and 35 cases were confirmed for canine demodicosis by skin scraping and amp; PCR examination.Result: The typical characteristics of Demodex spp. were confirmed in (20/35) 57.14% cases by skin scraping examination while PCR examination demonstrated (35/35) 100% by the amplification of an approximately 483 bp. Sequencing of PCR products were analyzed by BLAST and amp; the results indicated 99.7% identical to available sequences of D. canis MG372354 (1:99.7) and 98.8 identical with D. canis KU253790 (33:98.8) and amp; MG372359 (1:96.8). The sequence of the PCR product of positive samples was submitted to NCBI GenBank for accession number and MK177513 accession number was obtained for GenBank. From the present study it seems that Herbo-Nano medicine can be an effective alternative of Amitraz in case of demodicosis.
Sterile insect technique (SIT) is used, among other biological control tools, as a sustainable measure for the management of Ceratitis capitata Wiedemann (Diptera: Tephritidae) in many agricultural ...regions where this pest can trigger severe economic impacts. The tendency of wild females to remate multiple times has been deeply studied; it has been a common point of controversy when evaluating SIT programmes. Nevertheless, the remating potential of the released sterile males remains unknown. Here, under laboratory conditions, the remating capability of mass‐reared sterile males was determined. Wild‐type virgin females were offered to sterile males (Vienna‐8 strain), which had the opportunity to mate up to four consecutive times. The remating assays were carried out at 24 hr, 48 hr, 4 days and 7 days after the first mating. At the end of each tested time period, males were divided according to their mating response, mated or unmated, and subsequently reused for the next round of mating assays. The frequency of successful remating in each tested time period was obtained. Insemination was confirmed by determining the sperm transfer in mated female spermathecae by quantitative real‐time PCR. Our results demonstrate that 73% of the mass‐reared sterile males were able to remate 24 hr after the first mating, 55% of which remated again the day after. Close to 25% of the V8 sterile males tended to copulate in all of the four mating opportunities. The qPCR analysis of the spermathecae contents verified an effective transfer of V8 sperm to wild females with every mating; 99% of copulations resulted in sperm transfer. These findings shed light on the remating potential of V8 sterile males, an aspect until now underestimated in many SIT programmes.
Rhinosinusitis is a common disorder, influencing approximately 20% of the population at some time of their lives. It was recognized and reported with expanding recurrence over the past two decades ...worldwide. Undoubtedly, correct diagnosis of fungi in patients with fungal rhinosinusitis affects the treatment planning and prognosis of the patients. Identification of the causative agents using the standard mycological procedures remains difficult and time-consuming.
Based on clinical and radiological parameters, 106 patients suspected of fungal rhinosinusitis were investigated in this cross-sectional prospective study from April 2012 to March 2016 at an otorhinolaryngology department. In this study, internal transcribed spacer (ITS) and calmodulin (
) sequencing were respectively validated as reliable techniques for the identification of Mucorales and
to species level (both agents of fungal rhinosinusitis).
Of these, 63 (59.4%) patients were suspected of allergic fungal rhinosinusitis (AFRS), 40 (37.7%) patients suspected of acute invasive fungal rhinosinusitis (AIFRS), and 3 (2.8%) patients suspected of mycetoma. In patients suspected of AFRS, AIFRS, and mycetoma only 7, 29, and 1 had positive fungal culture, respectively. After ITS and
sequencing,
was the most common species isolated from non-invasive forms, and
and
were more frequently isolated from invasive forms.
is the most common agent of fungal rhinosinusitis in Iran, unlike most other reports from throughout the world stating that
is the most frequent causative agent of this disease.
The phylum Tardigrada consists of over 1,300 species that inhabit terrestrial, freshwater and marine environments throughout the world. In terrestrial habitats they live primarily in mosses, lichens, ...leaf litter and soil, whereas tardigrades in freshwater and marine environments are mainly found in sediments and on aquatic plants. More than 65 species have been previously reported in the state of Tennessee, USA.
Tardigrades present in moss cushions (
sp.) collected from a xerothermic habitat on the East Tennessee State University campus, Johnson City, TN, USA, were extracted, mounted on slides, identified, and counted. Additional samples of fresh dried moss were used for integrative analyses, including morphological analysis with phase contrast (PCM) and scanning electron microscopy (SEM), as well as molecular analyses of COI, 18S rRNA, 28S rRNA, and ITS-2 of the
and
species.
Five species were found, including two species new to science:
sp. nov. and
sp. nov.
sp. nov. differs from other members of the genus mainly by having a different type of dorsal cuticle and some other, more subtle, morphometric characters. In addition to the two new species,
and
were present, and males of
were found for the first time, the first record of males in the genus
.
sp. nov. is most similar to
, but it differs from
mainly by the stylet supports being situated in a more anterior position, shorter and narrower egg processes, and a smaller number of areoles around the egg processes. Moreover, the identification of
was confirmed as the first record for the USA by analysis of COI.
•The effects of GM plants on soil microbes were analyzed.•Foreign gene products released from residues of GM plants may affect soil microbes.•However, no consensus was reached on the effects of GM ...plants on soil microbes.
With the increase in the number of commercial applications and larger cultivation areas of genetically modified (GM) plants, their biosafety for soil microorganisms has become a controversial issue. The effects on the diversity and abundance of soil microorganisms are important components of evaluation of the biosafety risks of GM plants. So far, no definite conclusions have been drawn about whether GM plants can negatively affect soil microorganisms. In this review, we discuss the advances that have been made in recent years in the research into the effects of GM plants on soil microbial communities. It has been argued that foreign gene products that are released from the residue of GM plants into soil by root exudation may affect soil microbial communities. Moreover, foreign genes may change the genetic and functional properties of soil microorganisms via horizontal transfer. The advantages and disadvantages of various detection technologies—from classical culture-dependent methods to modern molecular protocols—are reviewed here. To accurately and comprehensively evaluate the effects of GM plants on microorganisms, we discuss the factors that should be considered in the assessment of risks of GM plants for soil microorganisms (e.g., foreign proteins, marker genes, plant varieties, and environmental factors), as well as the problems and prospects related to biosafety assessment platforms for GM plants.
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