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Cold mixed epoxy asphalt mixture (CMEAM) has excellent anti-deformation and fatigue performance which shows remarkable superiority on bridge pavements. Previous studies have done comprehensive researches on its properties through macroscopic experiments. However, few studies revealed the impact of gradation design on the road property of CMEAM through microscopic method. Thus, in this paper, X-Ray computed tomography (X-ray CT), nanoindentation (NI), field emission scanning electron microscopy (FESEM) and mercury intrusion porosimetry (MIP) are introduced to investigate the coarse aggregates contact degree, the thickness and elastic modulus of interfacial transition zone (ITZ) and the pore diameter distribution of CMEAM. Results showed that the contact points of coarse aggregates in EA-05 mixture are around 40 while those in EA-10 mixture are around 20. (40°, 50°) is the key inclination angle interval for EA-10 mixture type while (70°, 80°) and (10°, 20°) are of significance for EA-05 mixture type. The contact points and inclination angles are positively and inversely proportional to Marshall stability, respectively. In addition, the ITZ thickness of EA-05 and EA-10 mixture is below 10 μm and above 40 μm, respectively. The ITZ elastic modulus of EA-10 mixture is approximately 1.5 times than that of EA-05. The thickness and elastic modulus of ITZ are inversely proportional to low-temperature indirect tensile strain (εT). Meanwhile, EA-05 mixture has higher proportion in gel micropores and middle capillary pores while EA-10 mixture has higher proportion in mesopores and large capillary pores. Mesopores has the highest correlation coefficients with tensile strength ratio (TSR), followed by large capillary pores. The suggested proportion for above two type pores is around 21.5% and 51.0%, respectively.

The subject of the research was the oleogel structure of the OraMAF oral suspension, containing a mixture of active substances in which the oleogel environment is formed by purified olive oil in combination with soy lecithin (SL) and sorbitan tristearate (STS). The objective of the research was the development of an optimal methodology for the work process, enabling microscopic observation of the structure of the oleogel suspension. Purified olive oil structured with a combination of gelators SL and STS with the addition of a solid phase of fucoidan (F) and chondroitin sulfate (CS) powders was used for the study as an alternative to OraMAF suspension. The sedimentation method of separation of the medium and staining of solid phases of the suspension brought the expected results, allowing the observation of the network structure of the gel, which consisted of interlaced gelator fibers assembling into star formations. The most interesting structures were clearly the star-like structures created from the star-shaped microtubules trapping the CS and F solid particles, colored blue.

The use of insects and their processed animal proteins (PAPs) for animal nutrition creates the need for research into methods useful for routine surveillance for their presence. The aim of this study was to evaluate a modified microscopic method for the detection of particles of insects in poultry feed.

Background: Giardia intestinalis is one of the most common parasites in humans. Contaminated food and water can be a source of infection. Substances added to food are intended to increase its safety. We aimed to determination of the influence of various microorganisms and compounds that stimulate digestive functions, as well as preservatives and antioxidants on the detection of G. intestinalis by microscopic and immunoenzymatic methods. Methods: Twenty stool samples, archived in 1998-2018 in the Provincial Sanitary and Epidemiological Station in Bydgoszcz (Poland), collected both from patients referred for parasitic examinations by a doctor of a medical facility and from private individuals, were used to assess the impact of selected factors (such as bacterial strains, viruses and substances added to food) on the detection of G. intestinalis by microscopic and immunoenzymatic methods. Results: G. intestinalis was detected by both microscopic and immunoenzymatic methods with the same sensitivity (100%). The result of the G. intestinalis determination was positive in 90% of the samples after the addition of potassium sorbate, and in 25% of the samples after the addition of citric acid. Conclusion: The presence of other microorganism such as bacteria and viruses does not influence on the detection of G. intestinalis by microscopic and immunoenzymatic methods in stool samples. Citric acid as an antioxidant added to foods affects the detection of G. intestinalis. Due to the small number of samples used, it is necessary to continue research on the impact of various factors on the detection of protozoa.

Sarcocystis is one of the most important protozoa belonging to the phylum Apicomplexa, which is common among warm-blooded animals in all parts of the world, some of which are of zoonotic importance. Sarcocystis cruzi, S. bovifelis, and S. hominis are recognized in cattle. Due to the high occurrence of Sarcocystis in cow carcasses slaughtered in Iran, this study was conducted to investigate the contamination of Sarcocystis in beef slaughtered in Urmia industrial slaughterhouse using PCR. Also, the efficiency of the PCR method was compared with macroscopic and digestive (microscopic) methods. For this, a total of 80 esophageal and tongue samples, obtained from 40 carcasses was assayed. The Sarcocystis DNA was extracted according to the instructions of the Qiagen kit and the 18sRNA gene fragment was used using specific primers. The DNA product was digested with restriction enzymes, and their fracture pattern was evaluated. Out of 40 carcasses, in macroscopic and microscopic methods 2.5% and 72.5% were found positive for Sarcocystis, respectively. Besides, 36 cows were reported positive in terms of PCR contamination, which is 90% of the total samples. The results showed that the efficiency of PCR in detecting Sarcocystis is higher than the other two methods (p < /em>< 0.05) than the level of esophageal contamination. It was concluded that for efficient detection of Sarcocystis in cattle carcass, PCR can be applied along with the conventional methods.

Giardia intestinalis is one of the most common food-borne protozoa. The sensitivity of pathogens to physical and chemical factors is the basis for developing measures to reduce the incidence of the population. Several methods are available to detect the presence of G. intestinalis. The study determines the influence of 22 selected factors on the survival assessment and detection of G. intestinalis DNA in trophozoites axenically cultured. The influence of a given factor on the test result was observed in the case of 17 factors (77.3%) in the microscopic method, while only in the case of 3 (13.6%) substances in the real-time PCR method. Prevention of G. intestinalis infections, e.g., by ensuring food and water safety, is a crucial issue affecting public health. The experiment was conducted on trophozoites as the first approach. It is necessary to continue research and observe the epidemiological situation. In future studies, the impact of the studied factors on the survival assessment and detection of Giardia intestinalis DNA in axenically cultured cysts should be determined.

Abstract Background and aims Diagnosis of thrombotic microangiopathy (TMA) relies on microscopic schistocyte determination by an experienced microscopist. In addition, schistocytes can be found in non‐TMA–related disorders such as thalassaemia. We aimed to compare the accuracy of the automated haematology analyser Sysmex XN‐3000 for schistocyte detection, to that of the microscopy approach, in patients suspected of having schistocytosis. Methods Consecutive blood samples were collected between April 2016 and March 2017 at Siriraj Hospital, Mahidol University, Bangkok, Thailand. Specimens were collected from adults with suspected TMA or with thalassaemia trait and/or disease. All blood samples were examined by both microscopy and the analyser. Samples were considered to be positive for schistocytes (ie, schistocytosis) if they had a schistocyte count ≥1% by microscopy. The analyser's ability to determine schistocytosis was assessed by receiver operating characteristic (ROC) curve. Sensitivity, specificity, positive (PPV), and negative predictive value (NPV) of an appropriate cut‐off point were calculated, with manual microscopy as the standard. Quantitative agreement in schistocyte counts between the two approaches was assessed using 95% limits of agreement, Bland‐Altman plots, intraclass correlation coefficient, and concordance correlation coefficient. Results Ninety‐seven blood samples (62 suspected TMA and 35 thalassaemia) were collected. ROC curve analysis of the analyser for determining schistocytosis showed an area under the curve of 0.803 (95% confidence interval, 0.689‐0.917, P < 0.001). A cut‐off point of 0.6% yielded 86.1% sensitivity, 77.8% specificity, 94.4% PPV, and 56.0% NPV. The automated schistocyte count did not quantitatively agree with schistocyte counts by microscopy, neither in all blood specimens (mean of difference: −1.09; 95% limits of agreement, −11.9 to 9.7) nor in the subgroups (TMA, −0.88; 95% limits of agreement, −6.60 to 4.84; thalassaemia, −2.4; 95% limits of agreement, −14.10 to 9.30). The differences in the estimation of fragmented red blood cells between the methods tended to increase at higher schistocyte counts. Conclusion Sysmex XN‐3000 can be used for qualitative measurement of schistocytosis, but should not be used as a quantitative tool for schistocyte counting. Improvements are needed before this analyser's schistocyte detection feature can be recommended for use in clinical practice.

The official control of animal proteins in feed is focused on the prevention of Bovine Spongiform Encephalopathy (mad cow disease). The current legislation of the European Union is planned to avoid the feeding of animal by-products to the same species as its origin (ban of cannibalism, or species-to-species ban). With respect to the official control, the circumscription of the term species in legislation should be defined, and species-specific markers should be available. Markers will include primer sets, antibodies, near-infrared profiles or visual characteristics. The method of classical light microscopy is currently the only accepted method in the framework of the official detection of animal proteins. Besides the necessary development of complementary methods, either as stand alone methods or in combination, the visual characteristics used for a microscopic examination of meat and bone meal particles should be fully explored. Multivariate analysis of a range of characteristics of lacunae in bone fragments revealed that discrimination is possible between mammalian and avian bone fragments. Translation to features for every day practical use should be carried out very carefully, and only comprehensively collected information on a range of features will give a first indication of the source. Characteristics of hairs and feather filaments can be used to identify the origin of animal particles. An in situ identification method has been developed for antibody conjugation with troponin I in muscle fibers on a microscopic slide. A proof of principle is presented. Interlaboratory transferability and validation have still to be achieved. The development and testing of light microscopy markers in the framework of the SAFEED-PAP project revealed that a fine tuning of existing microscopic characteristics appears to be possible.

The TSE Roadmap published in 2005 (DG for Health and Consumer Protection, 2005) suggests that short and medium term (2005-2009) amendments to control BSE policy should include “a relaxation of certain measures of the current total feed ban when certain conditions are met”. The same document noted “the starting point when revising the current feed ban provisions should be risk-based but at the same time taking into account the control tools in place to evaluate and ensure the proper implementation of this feed ban”. The clear implication is that adequate analytical methods to detect constituents of animal origin in feedstuffs are required. The official analytical method for the detection of constituents of animal origin in feedstuffs is the microscopic examination technique as described in Commission Directive 2003/126/EC of 23 December 2003 [OJ L 339, 24.12.2003, 78]. Although the microscopic method is usually able to distinguish fish from land animal material, it is often unable to distinguish between different terrestrial animals. Fulfillments of the requirements of Regulation 1774/2002/EC laying down health rules concerning animal by-products not intended for human consumption, clearly implies that it must be possible to identify the origin animal materials, at higher taxonomic levels than in the past. Thus improvements in all methods of detecting constituents of animal origin are required, including the microscopic method. This article will examine the problem of meat and bone meal in animal feeds, and the use of microscopic methods in association with computer image analysis to identify the source species of these feedstuff contaminants. Image processing, integrated with morphometric measurements can provide accurate and reliable results and can be a very useful aid to the analyst in the characterization, analysis and control of feedstuffs.

This study was made to determine the effect of storage temperature and storage time on somatic cell counts (SCCs) in buffalo milk samples from quarters with no signs of clinical mastitis. The critical limit of SCC in raw buffalo milk is around 400,000 cells/mL. SCCs were measured in 32 Murrah buffalo milk samples by direct microscopic method. Milk samples were grouped from high score (> 3.50×105 cells/mL) to low score (< 1.50×105 cells/mL) according to the SCC. Each milk sample was divided into 3 aliquots and stored at 3 different temperatures (4°C, 21°C and 37°C). The SCC was recorded every hour up to 4 h and every 2 h up to 12 h. Storage of buffalo milk with a high SCC for 4 h at 4°C and 2 h at both 21°C and 37°C, respectively, decreased the log10 value of SCC compared to fresh milk. The log 10 value of buffalo milk SCC was lower after 1 h of storage than that determined for fresh milk at any tested temperature in low SCC samples. The results sugges that irrespective of storage temperature, buffalo milk samples should not be stored for more than 2 h before measurement of SCC by direct microscopic method.

Beer is a hostile environment for most microorganisms, but some lactic acid bacteria can grow in this environment. This is primarily because these organisms have developed the ability to grow in the presence of hops. It has been speculated that hop resistance is inversely correlated to resistance against oxidation, and this would have great impact on the use of various disinfectants in the brewing industry. In this study, we cultivated bacteria under aerobic and anaerobic conditions, and then investigated the in situ outgrowth of individual cells into microcolonies on MRS agar after exposure to the oxidizing agent peracetic acid (PAA). An automated microscope stage allowed us to analyse a much larger number of cells over extended periods of incubation. After PAA treatment, the lag time increased markedly, and extensive variation in morphology, µmax as well as stress resistance was observed between and within the tested L. brevis strains. The results suggest that aerobic cultivation increased the oxidative stress tolerance in L. brevis. The results also show that dead cells are randomly distributed in a microcolony and the majority of non-growing individual cells do not stain with a membrane impermanent dye (Propidium iodide), which indicates that PAA may not destroy the plasma membrane. In conclusion, the developed microscopic analysis of individual cells on MRS agar can provides faster results and more details of cell physiology compared to the traditional CFU method.

The aim of this study was to evaluate the potential of image analysis measurements in combination with the official analytical method (Commission directive 2003/126/EC) for the detection of constituents of animal origin in feedstuffs, in distinguishing between land animals (i.e. poultry vs mammals). Results obtained in the present study indicated that even though most of the variables measured were significantly different between mammalian and poultry in terms of mean, only a few can be used in distinguishing avian vs mammalian material. In conclusion, data presented indicate that some of the variables/descriptors provided by image analysis related to lacunae dimensions and features appear promising for a reliable distinction between animal meal at the level of vertebrate classes, while for further characterization, at higher taxonomic level, contribution of morphological variables deserves further investigation.

Background and Objectives: In Iran, anaplasmosis is normally diagnosed with traditional Giemsa staining method. This is not applicable for identification of the carrier animals. The aim of this study was to compare the detection of Anaplasma marginale in two different numbers of microscopic fields (50 and 100) using conventional Giemsa staining method compared with the PCR-RFLP technique. Materials and Methods: In this study, examinations were performed on 150 blood samples from cattle without clinical signs. Sensitivity and specificity of two microscopic fields (50 and 100 fields) were compared with A. marginale specific PCR-RFLP. The degree of agreement between PCR-RFLP and the two microscopic tests was determined by Kappa (κ) values with 95% confidence intervals. Results: PCR-RFLP showed that 58 samples were A. marginale, while routine microscopy showed erythrocytes harboring Anaplasma like structures in 16 and 75 blood samples determined in 50 and 100 microscopic fields respectively. Examination of 50 and 100 microscopic fields showed 25.8% and 91.4% sensitivity and 99% and 76.1% specificity compared to 100% sensitivity and specificity by PCR-RFLP. The Kappa coefficient between PCR-RFLP and Microscopy (50 fields) indicated a fair level of agreement (0.29). The Kappa coefficient between PCR-RFLP and Microscopy (100 fields) indicated a good level of agreement (0.64) Conclusion: Our results showed that the microscopic examination remains the convenient technique for day-to-day diagnosis of clinical cases in the laboratory but for the detection of carrier animal with low bacteremia, microscopy with 100 fields is preferable to Microscopy with 50 fields and molecular methods such as PCR-RFLP can be used as a safe method for identifying cattle persistently infected with A. marginale.