Age estimation with cemental incremental lines in normal and periodontally diseased teeth using phase contrast microscope: an original research

Background Age estimation is an important factor in forensic science for human identification. Teeth are considered to play a vital role as they resist decomposition at death unlike other tissues. This resistance and the gradual structural changes that take place throughout the life of an individual have made teeth useful indicators for age estimation. Dental cementum shows continuous apposition throughout the life of an individual. Tooth cemental annulation is a microscopic method for the determination of an individual’s age based on the analysis of incremental lines of cementum. Light microscopy as well as specialized microscopic methods have been employed to enhance the assessment of the cemental annulations. Periodontal disease is the most common dental problem affecting millions of people. Assessing the efficiency of the tooth cemental annulations method in periodontally diseased teeth is an important requisite. This study aims at assessing and evaluating the tooth cemental annulations in normal and periodontally diseased teeth using phase contrast microscopic method for age determination. Results A total of 60 teeth were included in the study and out of which 30 teeth were normal (sound teeth without any associated pathologies) and 30 were periodontally involved teeth respectively. Longitudinal ground sections were prepared and observed under phase contrast microscope. Measurements were made using an image analyzer software. The total width of the cementum was divided by the distance between two incremental lines. The eruption age of the tooth was then added to this to obtain the chronologic age for each individual. The results in the present study showed that tooth cemental annulations are applicable to periodontally sound teeth as well as in periodontally diseased teeth. There was no significant difference of estimated age from the actual age in both periodontally sound and periodontally diseased teeth. Normal teeth showed a reliability value of 92% and periodontally compromised teeth showed 96% respectively. There was no substantial influence of periodontal health on the estimated age. Conclusions The study concludes that the use of phase contrast microscopy in conjunction with image enhancement procedures improves the accuracy of age estimation and may serve as a reliable aid in forensic identification.

723. Cryptosporidium Detection in Preserved Stool Specimens: A Comparison Study of EIA, DFA, and Direct Microscopic Method

Background Cryptosporidium is an intestinal parasite that may cause diarrhea. Laboratory diagnosis largely relies on microscopic or immunology-based antigen detection. Direct fluorescent antibody (DFA) is considered the gold standard. Enzyme immunoassay (EIA) is an attractive alternative, but direct comparison studies for the performance together with the impact from different specimen preservation media are limiting. Methods We compared these three methods for the detection of Cryptosporidium oocysts (direct microscopic) or antigen (DFA or EIA) from stool samples preserved in either 10% buffered formalin, Cary-Blair/C&S, or Total Fix (MCC, Torrance, CA). The DFA from Meridian Bioscience (Cincinnati, OH) and the EIA using CRYPTOSPORIDIUM II (TechLab®, Blacksburg, VA) were performed according to the manufacturer’s instructions. The direct microscopic method was performed according to laboratory protocols, including direct wet mount, modified acid-fast stain, or permanent trichrome stain. Results A total of 140 samples, including 116 clinical specimens, 20 validation panel samples and 4 proficiency survey specimens, were examined (Table 1). The DFA and EIA methods produced 100% concordant results using all three preservatives, while the microscopic method had decreased sensitivity. All microscopic positives remained positive for both the DFA and EIA. Cross-reactivity from other parasites, such as Giardia, of the two immunoassays was not observed. Conclusion While the two immunological methods both outperformed the microscopic method, the EIA has the advantages of being objective, simple to perform, has less hands-on time, and thus makes it an attractive option for high throughput Cryptosporidium detection. Disclosures Kileen L. Shier, PhD, D(ABMM), MLS(ASCP)CM, Quest Diagnostics (Employee)

Research of efficiency of microsporia diagnostics methods

Objective — to investigate the effectiveness of different methods of microsporia diagnostics in children. Materials and methods. 50 children aged 2 to 16 (24 boys and 26 girls) were under survey. Depending on the clinical course, diagnosis and research results, all patients were divided into two groups: the 1st group included 40 children with microsporia (19 — with smooth skin microsporia, 13 — with scalp microsporia, 8 — with scalp and smooth skin microsporia); and the 2nd group consisted of 10 children in whom microsporia was not detected. The clinical diagnosis of all patients of the 1st group was confirmed by the results of PCR, microscopic, cultural and luminescent studies. The material for the study was scales from the smooth skin and scalp, as well as hair from the scalp of patients. 10 patients of the 2nd group did not have any clinical manifestations of microsporia and the results of the studies were negative. Results and discussion. The study with PCR in children with microsporia had 100 % positive result. Microsporum canis DNA was detected in all 40 patients. The microscopic method of the study was positive in 95 %. Bacteriological research revealed Microsporum canis in 85 %, while in 15 % the result was negative. Luminescent glow of hair in the rays of the Wood lamp in our study was observed in 87.5 % patients, while in 12.5 % it was absent. Conclusions. The study found that the most effective and accurate method is PCR. This is a method of modern accurate specific diagnostics of microsporia which allows the identification of the pathogen of Micro­sporum canis at the DNA level. Microscopic, cultural and luminescent research methods can also be used to diagnose this disease.

The Microscopic Detection of Animal Proteins in Animal Feed Regarding Bovine Spongiform Encephalopathy

Due to the actuality of spongiform encephalopathies and their proven spreading by means of animal feed containing meat and bone meal, the description and measurement of osteocytic lacunae contributes to more easily distinguish bone fragments in meat and bone meal. Transmissible spongiform encephalopathies (TSEs) have attracted a lot of attention, especially after 1986, when the first case of BSE (bovine spongiform encephalopathy) was detected. Since the outbreak of spongiform encephalopathy (BSE), the use of animal protein including bone meal as an ingredient in animal feed has been controlled by several regulations including Regulation (EC) 999/2001, Regulation (EC) 1774/2002, and Regulation (EC) 1234/2003. The classical microscopic method is the only official method for detecting animal protein in animal feed in the European Union (Commission Regulation (EC) 152/2009). By applying the microscopic method to the animal feed samples, we performed detection in order to determine the presence of animal proteins that originate from mammals and fish. The microscopic analysis of all 421 samples, of which 115 were raw materials for the production of animal feed, 230 were concentrates for ruminant nutrition and 76 were concentrates for non-ruminant nutrition (32 concentrates for laying hens and 44 concentrates for pigs), did not provide positive results, that is, no remains of animal tissues of mammalian origin were found in any specimen. Whereas in 10 out of 32 (31.25%) concentrates intended for non-ruminant nutrition (laying hens), pieces of fish tissue were found. In these samples, we usually detected the presence of fish bones, gills and scales.

THROMBOCYTOPENIA AND ELEVATED ALANINE AMINOTRANSFERASE LEVELS IN MALARIA PATIENTS

Objective: To study thrombocytopenia and elevated ALT levels in malaria patients reporting to Pak Med Level II+ Hospital XII and XIII, UN Mission Liberia, West Africa. Study Design: Cross sectional study. Place and Duration of Study: Departments of Pathology and Medicine, Pak Med Level II+ Hospital XII and XIII, UN Mission Liberia, West Africa, from Feb 2015 to Dec 2016. Methodology: In total of 100 febrile patients of both gender and all ages reported to Pak Med Level II+ Hospital XII and XIII, Liberia with clinical features of malaria and having positive malarial parasite (MP) on any of the methods of immune chromatography (ICT) or microscopic film were included. The febrile patients with typical clinical features of malaria but having negative MP both on ICT and microscopic methods were excluded.Pretreatment whole blood in EDTA was collected for testing MP on ICT and by microscopic method including MP index and platelets, while serum for ALT. Results: Out of total 100 patients, 75 were males, 25 females with mean age of 38 ± 5 years. MP was found positive on MP film in 95% and on ICT in 88% cases. Thrombocytopenia was found in 69% and ALT was elevated in 60% cases. Mean and SD for platelets count was 129 ± 72 x 109/L and of ALT 76 ± 66 U/L. Mean of MP index was 0.49 ranging from 0 to 3.5%. Correlation of MP index with platelets and ALT was found insignificant, although in malaria patients, platelets and ALT was inversely correlated significantly with each other showing low platelets along with elevated ALT levels. Conclusion: In malaria patients, thrombocytopenia and elevated ALT are frequent findings, which may not definitely correlate with MP index. So it is advisable to test for platelet count and ALT in all patients of malaria for early diagnosis and better management.

Karakterisasi parasit pada komoditas perikanan di Balai Karantina Ikan, Pengendalian Mutu, dan Keamanan Hasil Perikanan Semarang, Jawa Tengah.

Parasite is one of the infectious disease pathogens in fish which can cause secondary infection by bacteria, fungi and viruses. Therefore, fish quarantine activities are needed to prevent disease transmission and are carried out by the fish quarantine center. This study aimed to characterize the types of parasites that infect fish commodities that will be trafficked at the Fish Quarantine Center, Quality Control and Safety of Fishery Products in Semarang, Central Java. Parasitic examination was carried out using the original microscopic method by scraping the mucus, gills and internal organs, then characterizing it morphologically. The results showed that the infected parasites in fish samples included ectoparasites (Trichodina, Gyrodactylus, Dactylogyrus, Chilodonella, and Argulus) and endoparasites (Anisakis).Keywords:ParasitesFishery CommodityBKIPMHP SemarangABSTRAKParasit merupakan salah satu patogen penyebab penyakit infeksius yang dapat memicu terjadinya infeksi sekunder oleh bakteri, jamur maupun virus. Oleh sebab itu, perlu dilakukan tindakan karantina ikan untuk mencegah terjadinya penyebaran penyakit yang dilakukan oleh balai karantina ikan. Penelitian ini bertujuan untuk mengkarakterisasi jenis parasit yang menginfeksi komoditas ikan yang akan dilalulintaskan di Balai Karantina Ikan, Pengendalian Mutu dan Keamanan Hasil Perikanan Semarang, Jawa Tengah. Metode penelitian yang digunakan yaitu pemeriksaan secara mikroskopis menggunakan metode natif/langsung pada lendir, insang dan organ dalam ikan, dilakukan karakterisasi secara morfologis. Hasil penelitian menunjukkan bahwa sampel ikan terinfeksi parasit dari golongan ektoparasit (Trichodina, Gyrodactylus, Dactylogyrus, Chilodonella, and Argulus) dan endoparasit (Anisakis).Kata kunci:ParasitKomoditas Perikanan BKIPMHP Semarang

THE USE OF TOMATO POWDER IN PRODUCTION OF MAYONNAISE

It has been studied how tomato powder can be used in the production of mayonnaise. The content of essential amino acids in tomato powder has been compared with the FAO/WHO norms. Fresh plum tomatoes contain 0.158g of non-essential amino acids (in terms of 100g of dry matter), which covers 4.37% of the body’s requirements according to the standardised values approved by FAO/WHO. Tomato powder contains 0.14g of non-essential amino acids. The amount of essential amino acids in fresh tomatoes is 0.216g per 100g, and in powder, it is 0.181g per 100g. The amino acids that determine the intensity of sweetness have been established to amount to 0.165g in 100g of fresh tomatoes and to 0.116g in 100g of powder. So, in the course of drying, the product’s taste qualities related to feeling sweetness are reduced. It has been determined that the organoleptic properties of a product can be improved by adding tomato powder in the amount 1.8–2.2% and using a blend of oils. The mayonnaise samples obtained were cream-coloured with red particles of tomato powder. The samples had a soft structure and a more uniform and viscous texture than the control sample. The microscopic method has shown the homogeneous consistency of the product obtained. It has been noted that the absence of structure-forming agents does not reduce the quality indicators and does not impair the consistency of the finished product. According to the organoleptic parameters, the dose of tomato powder has been determined, which improves the taste of mayonnaise and does not make it oversweet. The research results show the prospects of using tomato powder not only as a carotene-containing raw material, but also as a raw material with a high content of amino acids. Besides, the use of tomato powder can modify the taste of such a product as mayonnaise.

Misidentification of Plasmodium ovale as Plasmodium vivax malaria by a microscopic method: a meta-analysis of confirmed P. ovale cases

Plasmodium ovale is a benign tertian malaria parasite that morphologically resembles Plasmodium vivax. P. ovale also shares similar tertian periodicity and can cause relapse in patients without a radical cure, making it easily misidentified as P. vivax in routine diagnosis. Therefore, its prevalence might be underreported worldwide. The present study aimed to quantify the prevalence of P. ovale misidentified as P. vivax malaria using data from studies reporting confirmed P. ovale cases by molecular methods. Studies reporting the misidentification of P. ovale as P. vivax malaria were identified from three databases, MEDLINE, Web of Science, and Scopus, without language restrictions, but the publication date was restricted to 1993 and 2020. The quality of the included studies was assessed using the Quality Assessment of Diagnostic Accuracy Studies (QUADAS). The random-effects model was used to estimate the pooled prevalence of the misidentification of P. ovale as P. vivax malaria by the microscopic method when compared to those with the reference polymerase chain reaction method. Subgroup analysis of participants was also performed to demonstrate the difference between imported and indigenous P. ovale cases. The heterogeneity of the included studies was assessed using Cochran's Q and I2 statistics. Publication bias across the included studies was assessed using the funnel plot and Egger’s test, and if required, contour-enhanced funnel plots were used to identify the source(s) of funnel plot asymmetry. Of 641 articles retrieved from databases, 22 articles met the eligibility criteria and were included in the present study. Of the 8,297 malaria-positive cases identified by the PCR method, 453 P. ovale cases were confirmed. The pooled prevalence of misidentification of P. ovale as P. vivax malaria by the microscopic method was 11% (95% CI: 7–14%, I2: 25.46%). Subgroup analysis of the participants demonstrated a higher prevalence of misidentification in indigenous cases (13%, 95% CI: 6–21%, I2: 27.8%) than in imported cases (10%, 95% CI: 6–14%, I2: 24.1%). The pooled prevalence of misidentification of P. vivax as P. ovale malaria by the microscopic method was 1%, without heterogeneity (95% CI: 0–3%, I2: 16.8%). PCR was more sensitive in identifying P. ovale cases than the microscopic method (p < 0.00001, OR: 2.76, 95% CI: 1.83–4.15, I2: 65%). Subgroup analysis of participants demonstrated the better performance of PCR in detecting P. ovale malaria in indigenous cases (p: 0.0009, OR: 6.92, 95% CI: 2.21–21.7%, I2: 68%) than in imported cases (p: 0.0004, OR: 2.15, 95% CI: 1.41–3.29%, I2: 63%). P. ovale infections misidentified as P. vivax malaria by the microscopic method were frequent and led to underreported P. ovale cases. The molecular identification of P. ovale malaria in endemic areas is needed because a higher rate of P. ovale misidentification was found in endemic or indigenous cases than in imported cases. In addition, updated courses, enhanced training, and refreshers for microscopic examinations, particularly for P. ovale identification, are necessary to improve the microscopic identification of Plasmodium species in rural health centres where PCR is unavailable.

Clustering energy calculation in light alpha-conjugated nuclei

In this paper, clustering energy was investigated in light alpha-conjugate nuclei considering cluster–cluster interaction instead of nucleon–nucleon interaction and using the phenomenological non-microscopic method. The observed energy levels were calculated from the rotational band of 8Be, 12C, and 16O isotopes. The results showed that these isotopes, in their cluster state, have a non-spherical structure.

Application of microscopy methods for evaluation of feed digestibility

The results of applying microscopic methods of analysis of pig feces to assess the digestibility of feed and diagnose various diseases of the gastrointestinal tract have been presented in the article. The microscopic method allows you to identify detritus, undigested fi ber, raw fat, fragments of animal feed ingredients, starch in the feces, gives an idea of the presence of foreign impurities (due to perverted appetite, contamination of feed), helminths, their eggs and other intestinal parasites. A number of preparations have been prepared to identify feed nutrients and feed components of the diet, and a number of coloring reagents were also used: Lugol’s solution for recognizing starch and its cleavage products (amylodextrin and erythrodextrin), Saathof’s reagent for detecting fat, and Hecht’s reagent for diff erentiating fat elements. The researches have been carried out under various magnifi cations of the microscope. Diff erent levels of detritus and undigested elements of feed in animals under the same conditions of rearing and feeding can identify individual features of digestion. The microscopic method of analysis does not require expensive equipment, reagents, and allows you to get the results of assessing the digestibility much faster than the methods of classical “wet” chemistry. This research method can be used by nutritionists to correct diets when using exogenous feed enzymes: phytase, xylanase, gluconase, amylase, mannanase, lipase, protease, etc., and provide results for discussion about the presence or absence of matrix eff ects from their use. The results of feces microscopy can give then idea of the quality of feed ingredients used: the content of non-starchy polysaccharides and the level of grain viscosity, the quality of animal ingredients (meat-bone, feather, fi sh meal), the level of indigestible fi ber in sunfl ower and soybean meals.