Search

Cystoisospora belli (C. belli) is the only pathogenic species of the Cystoisospora genus responsible for severe diarrhea in immunocompromised patients. Most common microscopic method of diagnosis is less sensitive due to intermittent shedding of oocysts. We developed a new single-run polymerase chain reaction (PCR)–based diagnostic assay for C. belli. A new single-run PCR-based diagnostic assay was standardized for the detection of C. belli. Diagnostic reproducibility and repeatability of the PCR assay were evaluated. A cross-sectional analytical study was done on a total of 354 stool samples collected from 331 immunocompromised patients with diarrhea. All the stool samples were tested for the presence of oocysts of C. belli and were also tested by our new PCR assay for C. belli. Three of the representative PCR products were confirmed by sequencing. Fisher’s exact test was used to compare the two proportions. Microscopy detected C. belli in 11/354 (3.1%) of stool samples, and the new PCR-based assay detected C. belli in 16/354 (4.5%). The new single-run PCR-based assay detected C. belli in all the stool samples which were tested positive by microscopy and additionally detected C. belli in five stool samples. The developed PCR assay detected statistically significant proportion of C. belli (p < 0.001) as compared to microscopy. The 795 base pair PCR product from one microscopy positive stool sample and two microscopy negative stool samples were confirmed by sequencing. Our newly developed single-run PCR-based detection assay for C. belli is robust and reproducible. It may be used for molecular diagnosis of cystoisosporiasis especially in transplant, pediatrics, and human immunodeficiency virus (HIV) positive patients.

The success of the anti-helminth mass treatment use Albendazole makes detection of soil transmitted helminth infections even more difficult to do microscopically. It is hoped that the molecular method was able to help increase the detectability of Soil Transmitted Helminth infection. The research aimed is to evaluate effectiveness of Albendazole administration in Bali, to identify the presence of β-tubulin gene as molecular diagnosis of STHs infection among children who treated by Albendazole. This study is a cross-sectional study that recruits elementary school children aged 6-12 years as subjects. Stool examination was carried out using the Kato-Katz technique, then followed by a molecular method using the B-tubulin gene as the target gene. The results showed that only 1 sample out of 140 examined using Kato-Katz was positive for Trichuris trichiura. 30 samples were then extracted from the faeces and performed Polymerase Chain Reaction. A total of 4 positive samples detected the B-tubulin Ascaris lumbricoides gene and 1 positive sample of the B-tubulin Trichuris trichiura gene. In conclusion, albendazole is still effective to treat STH infection, the molecular method has a higher detectability than the microscopic method.

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 2: 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.

Objective: This study aimed to inspect the occurrence of Haemoproteus tinnunculi (H. tinnunculi) in falcons from the central area of Saudi Arabia. Materials and Methods: Blood samples from 100 falcons species, including 55 Falco cherrug, 22 Falco peregrinus, 13 Falco pelegrinoides, and 10 Falco rusticolus, were collected from November 2018 to April 2019 and examined for H. tinnunculi by microscopic examination and nested PCR, targeting a cytochrome b (cytb) gene. Results: The prevalence was 1% by microscopic examination. The prevalence rate of H. tinnunculi was 1% by the microscopic method and 3% by PCR. Only F. cherrug was infected. In the sequence and phylogenetic analyses, the two cytb H. tinnunculi sequences were 100% identical and closely related to the Lithuanian isolate with 99.35% identity. Conclusions: This study presents the first report of molecular detection and characterization of H. tinnunculai in F. cherrug from the Kingdom of Saudi Arabia. [J Adv Vet Anim Res 2020; 7(4.000): 626-632]

Sarcocystis is considered as a common zoonotic coccidian parasite that infects intermediate hosts orally through ingestion of contaminated graze or water with protozoa oocyte. The purpose of study is to demonstrate the incidence of Sarcocystis infection in imported cattle in Duhok abattoir and achieve the gold conventional method for muscular tissue cyst and bradyzoites detection. Muscular tissue samples have been collected from esophagus, heart and diaphragm of 150 cattle. From a total of 1350 inspected samples (diaphragm, esophagus and heart) from three different imported origin have been 94%, 92% and 41.3% samples infected respectively as well as significant differences (p

Mycobacterium tuberculosis is a cause of tuberculosis (TB ) disease. In this research Mycobacterium tuberculosis can be detected using the Microscopic and Real-Time PCR. This research aims are to know the difference in the results of the examination Mycobacterium tuberculosis with microscopic method and Real-Time PCR. This research used an experimental research design and was tested using the Chi Square test. This results showed at significant difference (p=0.000) detection of pulmonary TB disease by microscopic and Real-Time PCR methods

When considering methods of detecting Cryptosporidium in patient samples, clinical and public health laboratories have historically relied primarily on microscopy. However, microscopy is time intensive and requires trained personnel to accurately identify pathogens that are present. Even with skilled analysts, the parasitemia level has the potential to fall below the level of detection. In addition, public health laboratories do not always receive specimens in fixatives that are compatible with the desired microscopic method. Antigen-based and molecular methods have proven to be effective at identifying Cryptosporidium at low levels and require less training and hands-on time. Here, we have developed and validated a real-time polymerase chain reaction (RT-PCR) laboratory-developed test (LDT) that identifies Cryptosporidium hominis and Cryptosporidium parvum, and also includes detection at the genus level to identify additional species that occasionally cause disease in humans. Results of the molecular test were compared with those obtained from modified acid-fast microscopy, immunofluorescent microscopy, an antigen-based detection rapid test, and a commercial gastrointestinal panel (GI panel). Of 40 positive samples, microscopy and antigen-based methods were able to detect Cryptosporidium in only 20 and 21 samples, respectively. The GI panel detected 33 of the 40 positive samples, even though not all specimens were received in the recommended preservative. The LDT detected Cryptosporidium in all 40 positive samples. When comparing each method for the detection of Cryptosporidium, our results indicate the LDT is an accurate, reliable, and cost-effective method for a clinical public health reference laboratory.

Issues related to the reproduction of animals have been and remain one of the most complex and relevant problems of biology and are constantly finding a direct and effective way out into livestock farming practice. The rational use of breeding sheep as producers is limited by the lack of standard, objective methods and methods for the timely assessment of their reproductive ability. The article presents the results of evaluating the sperm production of rams-producers. For the first time, the freshly obtained sperm of rams of Moldavian type producers of the Karakul breed was evaluated using the macroscopic method - ejaculate volume, color and smell, as well as the microscopic method - motility, sperm concentration in the ejaculate, total sperm count in the ejaculate sperm movements (VAP; VSL and VCL) as well as the percentage of abnormal forms of sperm. The experiments were carried out on sheep producers grown on a pedigree farm of the experimental farm of the Moldavian Scientific and Practical Institute of Biotechnology in Animal Husbandry and Veterinary Medicine. As a result of the studies, it was found that the ejaculate volume on average was 0.99 ± 0.04 ml, motility was 0.95 ± 0.02 and sperm concentration 1.51 ± 0.14 billion/ml. The percentage of pathological forms in sperm averaged 13.72 ± 0.61, an indicator that characterizes the high quality of sperm. The average quality indicator of ejaculates obtained from ram-producers of the Moldavian type of the Karakul breed corresponds to physiological standards for the Karakul breed. The average percentage of pathological forms of sperm found in ejaculates indicates that the rams were in good conditions of feeding and keeping. According to research, we can say that the Moldovan type of Karakul rams can be successfully used to obtain high quality ejaculates suitable for freezing sperm at 196°C.

Understanding drivers of fish maturity are essential to predict the productivity, stability, and resiliency of exploited populations. In terms of horse mackerel (Trachurus trachurus), in the eastern Atlantic Ocean and the Mediterranean Sea, size at maturity estimates date back to the 1940s and throughout its range. However, many older estimates may not reflect current maturity rates. This is because past methods were imprecise, or environmental or fishing effects may change maturity rates, and some stock areas have been overlooked. To address this data gap for the Central Mediterranean Sea, female horse mackerel size at maturity was estimated for two areas: the Strait of Sicily and the Tyrrhenian Sea. All ovaries were assigned a reproductive phase based on macroscopic methods, and the reliability of this method (94%) was validated with a microscopic method (i.e., gonad histology). Although the collected females exhibited similar condition, in terms of total body weight at a given length, as well as similar gonad-somatic indices, the size at median maturity in the Strait of Sicily was smaller (161 mm total length [TL]) than in the Tyrrhenian Sea (176 mm TL). Future sampling in a series of years with contrasting levels of productivity may help identify whether this is a broad latitudinal trend, or due to specific oceanographic drivers, such as estuarine outflow into the Tyrrhenian Sea or upwelling in the Strait of Sicily. Meanwhile, resource assessments should consider that these two areas represent two phenotypic stocks.

Determination of the stage of sexual maturation of an individual fish is commonly assessed using either microscopic or macroscopic staging. Microscopic staging examines either the internal cellular structure (histology) or the external appearance of the whole-oocyte (whole mount). Macroscopic staging is the visual evaluation of the external gonad development according to a standardised reference scale. Histology is the most widely used microscopic method and highly accurate but expensive and time consuming because it requires specialised equipment. Macroscopic staging is cheaper and faster but is associated with higher degree of error and bias. This study describe a methodological approach to assessing maturity stages of individual fish that is both inexpensive, rapid and more accurate than macroscopic staging. We have named it the gonometric method because it is based on direct measurements of the gonodasomatic index (GSI), which expresses gonad weight as a proportion of total or somatic weight. Immature, mature-active, and mature-inactive fish were classified using a logistic multinomial model giving a threshold value of GSI named here as GSIcut-off. The gonometric method was applied to four fish species that had different reproductive strategies, including demersal and pelagic species. For all four species the gonometric method estimated the maturity ogives with comparable precision to microscopic methods (p > 0.05). Across species, GSIcut-off values correlate with growth parameter of the von Bertalanffy growth function and length at 50% maturity, suggesting that the scaling of the GSIcut-off value is underpinned by fundamental allometric principles. Further testing of the gonometric method is required, but our results suggest that it has considerable potential for providing a faster and more accurate alternative to maturity staging without the need for extensive training requirements to ensure consistency across individual observers.

Blastocystis hominis is the most common intestinal parasite found in humans and many other hosts. Pathogenicity of Blastocystis spp. remains controversial, and it has been suggested that it may be associated with specific subtypes of the organism. This study identified the B. hominis subtypes and their prevalence rates in the northeast of Iran. A total of 1878 samples were collected from the northeast of Iran from January to December 2017. The patients’ demographic details were recorded. Samples were examined by a wet mount, and genomic DNA was extracted from positive samples. Also, PCR was done on the positive samples, and sequencing and phylogenetic analysis were subsequently performed. From 1878 collected stool samples, 152 (8.1%) Blastocystis samples were detected by the microscopic method. Of the 152 samples, Blastocystis spp. were found in 53.6% of the men and 28.9% of the women who showed clinical gastrointestinal symptoms, and a significant relationship was observed between gender and clinical symptoms ( P = 0.002 ). A meaningful relationship was found between the season and infection with this parasite ( P value = 0.003). The results of the sequencing of 22 PCR products showed the dominance of ST3, which was isolated from 10 (45.45%) patients, while ST1, ST2, and ST7 were found in 4 (18.19%), 7 (31.81%), and 1 (4.55%) patients, respectively. In this study, ST7 had a low prevalence in the northeast of Iran, and similar to previous studies, ST3 was the dominant subtype.

Trypanosomes are the causative agents of animal African trypanosomiasis (AAT) and human African trypanosomiasis (HAT), the former affecting domestic animals prevalent in Sub-Saharan Africa. The main species causing AAT in cattle are T. congolense, T. vivax, and T. b. brucei. Northern Uganda has been politically unstable with no form of vector control in place. The return of displaced inhabitants led to the restocking of cattle from AAT endemic areas. It was thus important to estimate the burden of trypanosomiasis in the region. This study was designed to compare the prevalence of animal African trypanosomes in cattle in Lira District using microscopy and polymerase chain reaction amplification (PCR) methods. In this cross-sectional study, a total of 254 cattle from the three villages of Acanakwo A, Barropok, and Acungkena in Lira District, Uganda, were selected by simple random sampling technique and screened for trypanosomiasis using microscopy and PCR methods. The prevalence of trypanosomiasis according to microscopic results was 5/254 (2.0%) as compared to 11/254 (4.3%) trypanosomiasis prevalence according to PCR analysis. The prevalence of trypanosomiasis infection in the animal studied is 11/254 (4.3%). Trypanosoma congolense was the most dominant trypanosome species with a proportion of 9/11 (81.8%), followed by T. vivax 1/11 (9.1%) and mixed infection of T. congolense/T. vivax1/11 (9.1%). Barropok village had the highest prevalence of trypanosomiasis with 6/11 (54.5%). There is a statistically significant relationship ( OR = 6.041 ; 95% CI: 1.634-22.331; p < 0.05 ) between abnormal PCV and trypanosome infection. Polymerase reaction amplification was the most reliable diagnostic method due to its high sensitivity and specificity as compared to the conventional microscopic method. Polymerase reaction amplification appears to have adequate accuracy to substitute the use of a microscope where facilities allow. This study, therefore, underscores the urgent need for local surveillance schemes more especially at the grassroots in Uganda to provide data for reference guideline development needed for the control of trypanosomiasis in Uganda.

Background: Among the human parasitic diseases, malaria is the main cause of morbidity and mortality. To prevent the high mortality and tracking malaria elimination efforts, a prompt and sensitive diagnosis is essential. This study aimed to compare High-Resolution Melting (HRM) and microscopic methods to diagnose Plasmodium falciparum and P. vivax. Methods: Eighty-one blood samples were collected from patients with clinical symptoms who were suspect to malaria in Chabahar district, southeastern Iran and also, from those who were referred to Malaria National Laboratory in the Tehran University of Medical Sciences, Tehran, Iran. Microscopic examination and HRM method were used to the diagnosis of Plasmodium parasites simultaneously. Results: Microscopic results revealed 45 positive cases (12 P. falciparum and 33 P. vivax) out of 81 collected samples while according to HRM analysis results 11 and 33 samples were identified as P. falciparum and P. vivax, respectively. HRM analysis also revealed 1 mixed infection of P. falciparum and P. malariae. Conclusion: HRM analysis provides a promising mean for simultaneous detection and discrimination of the Plasmodium spp. especially in mixed infection cases.

Microscopic examination results in patients with filariasis are often not identified by the presence of microfilaria, so it needs to be checked by Polymerase Chain Reaction (PCR). One PCR method uses High-Resolution Melting (HRM). The purpose of this study was to compare qPCR-HRM with microscopic examination methods to determine the types of microfilaria found in patients with filariasis. 19 samples were examined using a microscopic method and qPCR-HRM. The results of microscopic examination found no type of microfilaria and in qPCR-HRM identified B.malayi and W.bancrofti with peak temperature melting 78.1-78.7 ℃ and 80.2-80.8 ℃. The results of the study based on the comparison of two methods show that the types of microfilaria W.bancrofti and B.malayi can be found using qPCR-HRM.

This study applies the full velocity difference (FVD) model to investigate commuters’ driving process in a speed limit corridor with a toll station and explains how some factors affect the ...

Objective Herbal medicine products contribute a considerable part of the pharmaceutical market all over the world. Quality control of herbal medicines remains an obstacle in the herbal industry due to the existence of adulterants. Although some chemical methods are being employed to differentiate the herbal adulterants, they are time-consuming, costly and environmentally hazardous due to the use of dangerous chemicals. Hence, there is an urgent need to develop a rapid and reliable analytical method for the identification of herbal adulterants. Methods In the present study, the authors have employed different techniques such as Microscopy, Phytochemical screening, high-performance thin layer chromatography (HPTLC), gas chromatography coupled with mass spectroscopy (GC–MS) and near infra-red spectroscopy (NIRS) methods to differentiate Piper nigrum (black pepper) and its adulterant papaya seeds. Results Based on the results, microscopic method was found to be helpful to differentiate the herbal adulterants but required a lot of time and skilled human resource. Phytochemical methods are useful, but the process of extract preparation requiring the use of hazardous chemicals is not an environmentally friendly approach. The analytical techniques like HPTLC, GC–MS, and NIRS are useful methods, but require sophisticated instrumentation and skilled human resources involving time-consuming sample preparation steps. Conclusion In addition to conventional Botanical and Chemical characterization, NIRS was also found to be a very efficient, rapid and accurate technique for differentiating black pepper and papaya seed powders without the necessity for any sample preparation step.

This study aims to determine the type of endoparasitic worm Gnathostoma sp and the prevalence that infects rice field eel (Monopterus albus) with different grades. the method used is the descriptive method. Samples were taken from 3 different farm locations (Nor Aini, Banjar Regency, CV. Karya Bersama, Banjarbaru City, and Saberiannor Banjarbaru City), as many as 120 individuals with each grade (A, B, C, and D) totaling 30 individuals. Samples were examined using the microscopic method with 3 replications on each target organ, namely liver, meat, intestines, and kidneys. The results of the examination showed that Gnathostoma sp worm cysts were found, namely only in the liver, while in the intestines, flesh, and kidneys were not found. Based on the identification of Gnathostoma sp endoparasites seen from morphological characteristics, third-stage larvae (L3) in the phylum Nematoda, cylindrical body shape, transparent body color, and blackish brown, head shape resembling a light bulb, mouth, and has anus at the end of its body. Prevalence value in grade A = 40%; B= 30%; C= 40%, and D= 30%, this includes the category commonly or ordinary which describes that the parasite usually infects fish with a prevalence value range of 30-49%. It is still possible to send rice field eels outside of South Kalimantan Province, although the results of the examination of the test samples indicated the presence of the parasite infection Gnathostoma sp..

Des échantillons de matières fécales ont été prélevés sur 24 animaux apparte­nant à huit espèces différentes du parc zoologique de Hann au Sénégal. Ils ont été analysés par la technique de coproscopie microscopique qualitative de Tele­mann-Rivas et celle quantitative en cellules de Mac Master. Sur l’ensemble des échantillons, 66,7 % étaient parasités et chaque animal positif était infesté par au moins une espèce d’oeufs d’helminthes, alors que les protozoaires n’étaient présents que chez quatre animaux. Chez les carnivores, les oeufs de parasites, comme Toxascaris leonina et Toxocara cati, et ceux d’ankylostomidés étaient les plus présents, alors que chez les primates les oeufs de Trichuris sp. et Entamoeba sp. ont été identifiés. Cette étude constitue une base pour la mise en place de traitements chez ces animaux.

The present study conducted on thirty patients with scabies. Blood samples were collected before and after treatment in addition to 30 controls, that used in immunological study to determine the effect of infection with scabies on concentrations of IL-31, β-endorphin, PAR2 (Protease-activated receptor 2) before and after treatment. The sim of this study is to examine the immunological response in patients after becoming infected with scabies. All these cases were collected from Healthy centers of camps Kaparto1and 2, Sharyia and Khanki in Duhok governorate and diagnosed by clinical examination and light microscopic method for examining the Sarcoptes scabiei mite or its eggs, at laboratories of these centers from March 2019 to about 30 January 2020. Immunological parameter including interleukin-31, β-endorphin, PAR2 (Protease-activated receptor 2) were assessed using ELISA. Results of study showed that IL-31, β-endorphin, PAR2 (Protease-activated receptor 2) significantly increasing (P

The present study conducted on thirty male patients who were infected with scabies. Blood samples collected before and after treatment in addition to 30 male controls, were used in immunological study to determine the effect of infection with scabies on concentrations of Tropomyosin, Paramyosin, Melatonin, and Cathepsin-D before and after treatment. All these cases were collected from Healthy centers of camps Kaparto1and 2, Sharyia and Khanki in Duhok governorate and diagnosed by light microscopic method for examining the Sarcoptes scabiei mite or its eggs, at laboratories of these centers from March 2019 to about 30 January 2020. The results of study revealed the levels of Tropomyosin, Paramyosin, Cathepsin-D were significantly increasing (P

Crossbreeding with Duroc breed allows to improve meat quality, but no data is available regarding specific influence of Duroc breed on characteristics of meat in the case of crossbreeding with various breeds. The aim of the present research was to evaluate the effect of crossbreeding Polish Landrace dames with Duroc sires on quality features of meat in reference to Polish Landrace breed. The objects of the study were Longissimus dorsi lumborum pork muscles obtained from Polish Landrace breed and Polish Landrace x Duroc crossbreed animals. Sarcomere length measurements were conducted using microscopic method and basic chemical composition measurement was analyzed using spectrophotometric scanning. Texture analysis of meat samples, performed after thermal treatment was expressed by Warner-Bratzler shear force and color analysis was obtained using CIE L*a*b* color system. No differences in sarcomere length, shear force as well as components of color values between pork meat originated from Polish Landrace breed and Polish Landrace x Duroc crossbreed were observed. Analysis of basic chemical composition revealed higher fat and lower ash contents in the case of meat of Polish Landrace breed animals. It was concluded that the actual impact of breed on meat characteristics is possibly altered by other factors. It may be suggested that influence of basic chemical composition on color of meat is breed-related. Cruzamentos com a raça Duroc permite a melhoria da qualidade da carne suína. No entanto, não há dados disponíveis na literatura sobre a influência específica da raça Duroc sobre características da carne após cru zamento com outras raças. O objetivo do trabalho foi avaliar o efeito do cruzamento de matrizes Polish Lan drace com reprodutores Duroc sobre as características de carne de qualidade, em referência à própria raça Polish Landrace. O objeto do estudo foi o músculo dorsal lombar (Longissimus dorsi lumborum) obtidos de suínos das raça Polish Landrace breed e da raça mestiça entre Polish Landrace e Duroc. Medidas do comprimento do sarcômero foram realizadas por microscopia e a de terminação da composição química básica foi analisada por espectrofotometria. A análise de textura de amostras de carne, realizada após tratamento térmico, foi ex pressa pela força de cisalhamento Warner-Bratzler enquanto a análise de cor foi realizada pelo sistema de cores CIE L*a*b*. Não foram observadas diferenças no comprimento do sarcômero, força de cisalhamento e coloração entre as carnes provenientes das raças Polish Landrace e mestiça Polish Landrace x Duroc. A análise da composição química revelou maior teor de gordura e menor teor de cinzas na carne proveniente de animais da raça Polish Landrace. O trabalho permitiu concluir que o impacto da raça nas características da carne é possivelmente influenciado por outros fatores. No entanto, pode-se sugerir que a composição química básica no padrão de cor da carne está relacionada com a raça.

Background Food and water-borne illness caused by ingestion of (oo)cysts of Cryptosporidium and Giardia is one of the major health problems globally. Several methods are available to detect Giardia cyst and Cryptosporidium oocyst in food and water. Most of the available methods require a good laboratory facility and well-trained manpower and are therefore costly. There is a need of affordable and reliable method that can be easily implemented in resource limited settings. Methodology/Principle findings We developed a smartphone based microscopic assay method to screen (oo)cysts of Cryptosporidium and Giardia contamination of vegetable and water samples. The method consisting of a ball lens of 1 mm diameter, white LED as illumination source and Lugols's iodine staining provided magnification and contrast capable of distinguishing (oo)cysts of Cryptosporidium and Giardia. The analytical performance of the method was tested by spike recovery experiments. The spike recovery experiments performed on cabbage, carrot, cucumber, radish, tomatoes, and water resulted in 26.8±10.3, 40.1±8.5, 44.4±7.3, 47.6±11.3, 49.2 ±10.9, and 30.2±7.9% recovery for Cryptosporidium, respectively and 10.2±4.0, 14.1±7.3, 24.2±12.1, 23.2±13.7, 17.1±13.9, and 37.6±2.4% recovery for Giardia, respectively. The spike recovery results are comparable with data obtained using commercial brightfield and fluorescence microscope methods. Finally, we tested the smartphone microscope system for detecting (oo)cysts on 7 types of vegetable (n = 196) and river water (n = 18) samples. Forty-two percent vegetable and thirty-nine percent water samples were found to be contaminated with Cryptosporidium oocyst. Similarly, thirty-one percent vegetable and thirty-three percent water samples were contaminated with Giardia cyst. Conclusions The newly developed smartphone microscopic method showed comparable performance to commercial microscopic methods. The new method can be a low-cost and easy to implement alternative method for simultaneous detection of (oo)cysts in vegetable and water samples in resource limited settings., Author summary Food and water-borne illness arising from the consumption of contaminated food and water are serious health hazards globally. Cryptosporidium and Giardia are the major food and water‒borne parasites. The infection occurs mainly by (oo)cyst phase of the parasites. People in developing countries are more vulnerable to these parasites where infection is more likely underdiagnosed and underreported due to limited resources for detection. There is need of a method that is affordable and easy to implement. In this study, we developed and optimized a novel smartphone microscope method that can detect and quantify the (oo)cyst of the parasites in food and water samples. The developed method is easy to implement and affordable and provides similar performance to the other commercially available microscopic methods.

Background: Multiplex PCR examination is one of the molecular examination methodologies applied to detect soil-transmitted helminth (STH) infection. Optimization of the multiplex PCR method is a complex process, but it is necessary to obtain both a correct detection process and a satisfactory DNA product. Objective: To determine whether multiplex PCR can be optimized to diagnose STH from Indonesian isolates, and to find the optimal method for detection of A. lumbricoides, T. trichiura, and N. americanus infections in stool that have been stored for 3 years. Methods: A total of 15 samples were examined, and these samples were previously examined by using a microscopic method, then continued with optimization steps. Result: The optimal PCR mixture used primers targeting COI gene for A. lumbricoides, 18S rDNA for T. trichiura and ITS1 for N. americanus, 15 µl of Go Taq Green Master Mix, 5 µl of the 3 pairs of primers, 5 µl of DNA template and 4 µl of DdH2O, and the condition was 30 minutes of 950C denaturation, 30 second of 530C annealing and 1 minute of 720C extension, repeated for 35 cycles. Conclusion: Multiplex PCR can be optimized for STH detection from Indonesian isolates. The successful detection using the multiplex PCR method was influenced by sample preparation prior to DNA isolation, which includes several steps i.e. homogenization of samples using bead beaters and passing samples on liquid nitrogen rapidly.

As the largest inland sea of China, the Bohai Sea has confronted significant environmental changes in recent decades, and a shift of phytoplankton community from diatoms to dinoflagellates was observed by microscopic method in previous studies. However, the counting of some small-sized non-diatoms (e.g., flagellates and cyanobacteria) have long been neglected due to the limitation of this method. In this study, we measured the phytoplankton community structure and abundance in the central Bohai Sea in summer and autumn of 2015 by employing both microscopic method and high-performance liquid chromatography (HPLC) pigments - CHEMTAX analysis. The results of microscopic counting showed that the phytoplankton community was characterized by diatoms, dinoflagellates, and chrysophytes, which contained a total of 107 species. Dinoflagellates and diatoms co-dominated in summer while the latter became the dominant group in autumn. Compared with microscopic counting, CHEMTAX analysis allowed identification of a larger number of phytoplankton groups (including 8 phyla). The results of CHEMTAX analysis showed that besides diatoms and dinoflagellates, the small-sized non-diatoms (i.e., flagellates and cyanobacteria) also played an important role. Diatoms and cyanobacteria dominated the total Chl a (contributing 32.5% and 17.1%, respectively), followed by cryptophytes, dinoflagellates, and prasinophytes in the summer. A short-term summer to autumn variability of phytoplankton abundance and community was observed. Temperature and nutrient availability affected the succession of the phytoplankton community from diatoms and non-diatoms co-dominance in the summer to absolute diatoms dominance in the autumn. A direct comparison of algal cell abundance and CHEMTAX biomass showed that a significant positive correlation was observed only for dinoflagellates (P 0.01) in the autumn. Our work provides some basic information on the dynamics of small-sized flagellates and cyanobacteria in this area, and also suggests that a combination of both the methods is needed in future investigations to better understand the current status as well as the future changes of phytoplankton community.

Background/aims: Ankaferd blood stopper (ABS) is a mixture of certain ratios of 5 different plant roots (Thymus vulgaris, Glycyrrhiza glabra, Vitis vinifera, Alpinia officinarum, and Urtica dioica). The aim of this study is to evaluate the histopathological effects of ABS on articular cartilage in vitro. Materials and methods: Twenty-one albino Sprague Dawley rats were randomly allocated to 3 groups: 0.1 mL of saline was injected in the first group, 0.1 mL of ABS was injected in the second group, and 0.1 mL of blood and 0.1 mL of ABS were injected in the third group. One month later all rats were sacrificed. Specimens were obtained for histopathological evaluation. Results: Significant results were detected in the groups with respect to International Cartilage Repair Society and synovial proliferation scores (P < 0.05 and P < 0.01). According to inflammatory cell infiltration and fibrin formation scores, there was no significant difference between group 1 and group 2 (P < 0.01), although there was significant difference between group 3 and the other groups (P > 0.05). Conclusion: ABS and hemarthrosis had toxic effects on knee cartilage. The side effects were increased with the combination of hemarthrosis and ABS. As a result, ABS had unexpected effects on experimental hemarthrosis.

The aim of the study was to gather and analyse available data concerning the presence of terrestrial animal constituents in feeds in Poland. A microscopic method of identification of the contaminants was used. Between 2012 and 2013, overall 16 139 samples of feeds were analysed, from which 282 (1.75%) contained elements from terrestrial animals. The percentage of feeds contaminated with such elements was lower in 2013. In 2012, among 8 499 samples analysed, 203 (2.39%) contained ingredients of animal origin, and in 2013, the elements were found in 79 (1.03%) out of 7640 samples. The percentage of feed samples positive for processed animal protein was relatively low and steadily decreasing. Furthermore, the microscopic method demonstrated to be a very sensitive technique for the detection of constituents of animal origin. Keywords: feeds, animal protein, contamination, microscopic method. Introduction Processed animal proteins (PAP) as well as meat and bone meal (MBM) of terrestrial animals are banned for use in animal feed in European Union (6, 9). The ban was implemented to eliminate the risk of TSE (transmissible spongiform encephalopathy) transmission in livestock and subsequent transmission to humans (2, 3, 22). The detection of animal proteins is based on the microscopic method (7). The current official description of the method is published in Commission Regulation (EU) No 51/2013 (8). The strength of the microscopic method is the combination of the relatively fast detection of animal proteins, sufficient detection of contamination levels as low as 0.02% (11, 24), indication of the type of the detected materials, insensibility to sterilisation temperature, and well established distinction between prohibited (

Background. Sperm is very sensitive to changes in temperature, this can cause changing the characteristics of spermatozoa. To perform sperm analysis it is necessary to determine the optimal storage temperature to keep the characteristic of sperm samples remain representative. Objective. The purpose of this study to determine the effect of variation in storage temperature on sperm vitality, in order to determine the optimal storage temperature of sperm samples when delay of examination. Methods. In this study we used microscopic method with supravitalstainting using 0,5% eosin staint. Dead spermatozoa will be stained by eosin and live spermatozoa will not be stained by eosin.The study was conducted on 8 subject of the Department of Health Analyst Poltekkes Denpasar that selected by randomly simpling with One Group pretest-posttest design. The measured variable is the value of sperm vitality before and after treatment. Results : Analysis with T-Paired test showed significant difference before and after treatment (p

Introduction: The objective of the study was to comparatively evaluate the porosity distribution in the root canals obturated using the single-cone technique and thermoplasticized technique. Materials and Methods: A total of 42 permanent single-rooted human extracted teeth were divided equally into two groups based on the technique employed: single-cone technique and thermoplasticized technique. Following postobturation restoration, void volume was analyzed by cone-beam computed tomography (CBCT) and void location was evaluated by microscopic method at three levels: coronal, middle, and apical. Independent Student't-test and Chi-square test were used for analyzing the obtained results. Results: On comparing the void volumes as seen in CBCT images between the two groups, the thermoplasticized technique showed greater but insignificant void volume (P = 0.18). When the association of the location of the void with the void area percentage was analyzed between the two techniques, significant associations were seen in the middle (P = 0.016) and the apical levels (P = 0.031). Conclusion: The single-cone technique showed a comparable seal with the thermoplasticized technique. Voids were predominantly seen at the sealer–cone interface and sealer–root canal wall interface for single-cone technique and in between the condensed gutta-percha for thermoplasticized technique.

As parasitological or microscopic method is the gold standard and the best method for diagnosis of malaria, so fixed Geimsa-stained blood slides in the form of thick and thin blood smears are the most important data collections of malaria, especially historical slides. The parasites are dead but their DNA is valuable for many molecular biologic researches. A simple and efficient method for catching and extraction malaria parasites' DNA with a desired yield from dried and stained blood on slides is the first and major step. Introduction of an applicable, efficient and inexpensive DNA catching method and assessment of its performance in following molecular applications was the main objective of present study.

Moldavian dragonhead (Dracocephalum moldavica L.), due to its pleasant lemon scent and medical effects, has acquired increasing impact as functional food. The high diversity within the genus, limited supply not keeping pace with the growing demand, the morphological similarity with other Labiatae, and trading under the common name Turkish Melissa invite adulteration by surrogate species. We have developed several verified reference accessions of D. moldavica L. along with potential surrogate species to compare different approaches of authentication, also in commercial samples. We report on three strategies of authentication—a microscopic method, based on the relative size of epidermal pavement cells versus palisade cells, and two strategies of genetic authentication based on the barcoding marker large subunit of ribulose-1,5-bisphosphate carboxylase oxygenase (rbcL). We can detect single-nucleotide exchanges between D. moldavica L. and the potential surrogate species Melissa officinalis L. and Nepeta cataria L. by restriction fragment length polymorphism (RFLP), and we show that we can use this to verify the presence of D. moldavica even in dried and highly fragmented mixtures from commercial samples. We further develop a third strategy derived from the so-called amplification refractory mutation system (ARMS), based on multiplex PCR of the rbcL marker upon addition of specifically designed intermediate primers that will generate a diagnostic second band in case of D. moldavica L., but not for the surrogate species. We demonstrate that this ARMS approach is superior to the RFLP strategy, because it safeguards against experimental fluctuations, can unequivocally verify the presence of D. moldavica in commercial samples declaring its presence, and yields a clear outcome in a one-step protocol.

The article is devoted to research of functional and technological properties of powders from banana, carrots, strawberry, apple, spinach and orange received by cold spray drying. Expediency of vegetable and fruit powders use in the confectionery industry, including their addition during production of confectionery semi-finished products has been proved. Organoleptic properties of powders have been defined, namely: appearance and consistence, taste, smell and color, results are presented in the table. By the microscopic method dispersion of powders from banana, carrots, strawberry, apple, spinach, orange has been studied, results of researches are presented by charts. Also technological indicators of plant powders have been investigated, namely: the ability to bind moisture, water absorption coefficient, the emulsifying ability and ability to hold fat.

The Chinese Materia Medica, Tiepishihu, used as a tonic for over one thousand years, is a well-known precious medicine in China. According to the Chinese Pharmacopoeia, its source is the species Dendrobium officinale Kimura et Migo, which is distinguished from other species in Dendrobium genus. However, these species from the same genus are similar with Tiepishihu and caused confusion in the market. To find a quick and simple method to distinguish Tiepishihu from other similar species, histologic and microscopic methods were combined together to investigate the transverse section of stem of Tiepishihu and other similar species. Phloroglucinol test solution with hydrochloric acid was used to reveal the lignified tissue by staining the transverse section of Tiepishihu and similar species. Results revealed the unique identification characteristics to distinguish Tiepishihu from similar species, which were difficult to distinguish by other methods. The identification characteristics of Tiepishihu include the cells of vascular bundle sheath were stained red, parenchyma cells were not stained red. What's more, other species can be distinguished from each other with microscopic and histological characteristics. These characteristics proved stable and can be easily observed by normal light microscopic examination. This method is rapid, accurate, stable, and inexpensive.

In vitro susceptibility tests provide information on the intrinsic response of Plasmodium vivax to antimalarials, free from confounding factors such as host immunity or relapse. This study examined the utility of radioisotope and PicoGreen assays as alternatives to the traditional microscopic examination for assessing response of P. vivax to antimalarial drugs. There was no significant difference in the mean chloroquine IC50 of P. vivax (n = 40) as determined by the microscopic (33.4 ng/ml), isotopic (33.6 ng/ml), and PicoGreen (39.1 ng/ml) assays, respectively (F = 0.239, df = 2, 51, and p = 0.788). However measurement of IC50s by the microscopic method was slightly more successful in producing valid assays (57%), compared to the isotopic (32.5%) and PicoGreen (45.5%) methods. In a paired comparison of 20 fresh and cryopreserved isolates as examined by the microscopic method, there were no significant differences between the mean IC50 responses (T = 1.58, df = 15, and p = 0.34). Detailed methodologies for the short time culture of field and cryopreserved P. vivax are described. Although the microscopic in vitro assay provides a useful method for characterizing the drug susceptibility phenotype of P. vivax isolates, its utility is limited by a laborious methodology and need for highly skilled microscopists. Future efforts should focus on further development of high throughput assays such as the PicoGreen assay as described in this study.

Ex vivo antimalarial sensitivity testing in human malaria parasites has largely depended on microscopic determination of schizont maturation. While this microscopic method is sensitive, it suffers from poor precision and is laborious. The recent development of portable, low-cost cytometers has allowed us to develop and validate a simple, field-optimized protocol using SYBR green and dihydroethidium for the accurate and objective determination of antimalarial drug sensitivity in freshly isolated Plasmodium vivax and Plasmodium falciparum .

Background/aims: Epidermal thickness is frequently measured by light microscopy. The preservation and staining methods may alter the proportions of the specimens and thereby influence the measurements. The aim of the present study was to describe: 1) a standardized light microscopic method to quantify the thicknesses of the stratum corneum and the cellular epidermal layers, 2) the variations according to preparation and staining techniques, and 3) the observer variability. Methods: One hundred and sixty skin biopsies from 67 human volunteers were included. The cellular epidermis and the stratum corneum were estimated in sections preserved by freezing and subsequent preparation with cryostat or formalin-paraffin techniques. The slides were stained with haematoxylin-eosin or er-ythrocin, and thicknesses of the stratum corneum and the cellular epidermis were measured by a calibrated ruler and an ocular grid, respectively. Results: The formalin fixation gave slightly higher values for the cellular epidermis than the cryostat technique. In comparison to erythrocin staining, haematoxylin-eosin gave a significantly thinner stratum corneum. No significant inter- or intra-observer variation was found for the thickness of the stratum corneum assessed twice by two experienced observers. However, the two observers differed slightly from each other on the thickness of the cellular epidermis. Conclusion: It is found that thickness measurements of the stratum corneum and the cellular epidermis are reliably performed on cryostatic cut sections stained with haematoxylin-eosin.

The villous tree of the human placenta is a complex three-dimensional (3D) structure with branches and nodes at the feto-maternal border in the key area of gas and nutrient exchange. Recently we introduced a novel, computer-assisted 3D light microscopic method that enables 3D topological analysis of branching patterns of the human placental villous tree. In the present study we applied this novel method to the 3D architecture of peripheral villous trees of placentas from patients with intrauterine growth retardation (IUGR placentas), a severe obstetric syndrome. We found that the mean branching angle of branches in terminal positions of the villous trees was significantly different statistically between IUGR placentas and clinically normal placentas. Furthermore, the mean tortuosity of branches of villous trees in directly preterminal positions was significantly different statistically between IUGR placentas and clinically normal placentas. We show that these differences can be interpreted as consequences of morphological adaptation of villous trees between IUGR placentas and clinically normal placentas and may have important consequences for the understanding of the morphological correlates of the efficiency of the placental villous tree and their influence on fetal development.

Callophyllum inophyllum Linn. is one of potential tree species for commercial plantation as biofuel sources. This species was generally propagated by generative method and seldom vegetative method in mass planting stock production. A completely randomized design was arranged in factorial experiment to identify the effect of three grafting methods and four origins of scion in mother tree on the growth of grafted plants. The growth of grafted plants was observed at nursery and the development of graft union anatomy was observed through microscopic method in laboratory. The result showed that grafting method significantly influenced to shoot number and shoot length, with the best result showed by veneer graft method. Position of scion in mother trees influenced shooting rate and the best result was gained through the scion from the lower part of the tree crown. The survival rate was 72-100%, shooting rate was 68-100% with 1,9 shoots per grafted and average of shoot length was 0,65 cm.The growth of graft union showed earlier by callus formation and then differentiated to a vascular tissue

The purpose of this study was to investigate usefulness of the pine needles extract on Atopic Dermatitis(AD)-like skin lesions. To investigate the effect of pine needles extract in vivo, atopic dermatitis (NC/Nga) mice using DNCB (2.4-Dinitrochlorbenzene) was used. NC/Nga mice were challenged with DNCB during 2 weeks to develope AD-like skin lesion. After that, pine needles extract was applied to AD-like skin lesion on the backs of the NC/Nga mice during 3 weeks. The efficacy of pine needles extract in the NC/Nga mice was evaluated by measurement of the skin lesion severity(NC mouse score), the serum IgE level, epidermal thickness changes, and mast cell number. Blood was collected from the retro-orbital area and the level of IgE in the blood was measured. The epidermal thickness and mast cell number were observed by microscopic method after H&E stain. The serum IgE levels were decreased after treatment with pine needles extract. The epidermal thickness and mast cell number were decreased after treatment with pine needles extract. To conclude, the topical application of pine needles extract suppressed the progression of AD-like skin lesion.

Sophisticated modern research tools for evaluation of the plant drugs are available today but microscopic method is one of the simplest and cheapest methods to start with for establishing the correct identity of the source materials. the leaves of Terminalia catappa (combretaceae) are reported to have good medicinal values in traditional system of medicines. But the pharmacognostic and Phytochemical standardization of the leaves were not validated till date. With the aim of drawing the pharmacopoeial standards for this species, the present study deals with pharmacognostical parameters for the leaves of Terminalia catappa which mainly consists of Macroscopical and microscopical characters, physio-chemical constants, quantitative microscopy parameters and Preliminary phytochemical screening. this information will be of used for further pharmacological and instrumental evaluation of the species and will assist in standardization for quality, purity and sample identification.

Sputum smear microscopy is an easy, inexpensive, and rapid method for detecting tubercle bacilli when there are more than 10,000 bacilli/ml in the original sputum. Furthermore, because the microscopic method provides not only quantitative, but also qualitative information, such as the shape of bacilli, it has remained significant. We have previously developed and reported panel test slides made from polyacrylamide-based artificial sputum (PBAS) mixed with both cultured THP-1 cells and nonpathogenic mycobacteria. In this paper, we report an improved preparation method for PBAS for panel test slides that provides a simplified method and enhanced availability with high consistency in each grade and in which only negative PBAS is prepared from polyacrylamide and cultured THP-1 cells and mixed with graded formalin-fixed Mycobacterium tuberculosis solution (FFTBS) containing oral flora and Pseudomonas aeruginosa on the slides. In the smears prepared using this improved method, the numbers (average ± standard deviation [SD]) of acid-fast bacilli (AFB) in 300 fields (2- by 3-cm smear) in eight smears of each grade ranged from 5 to 9 (6.4 ± 1.4), from 59 to 88 (74.6 ± 10.0), from 503 to 912 (705.0 ± 145.7), and from 1,819 to 3,256 (2133.3 ± 478.0) in ±, +, ++, and +++ smears, respectively. In addition, this preparation method provided high similarity to the microscopic appearance of bacilli and background seen in the actual patient sputum, with high feasibility. These results revealed that our new PBAS had high authenticity in the appearance and consistency in each grade, which could make it valuable as a reliable artificial sputum for the training of microscopists.

Heritage potato varieties in Canada are historic old varieties collected from Canada and various countries before the formal establishment of the Canadian plant variety registration system and may be a valuable gene resource for beneficial traits in potato breeding and bioproducts development. Greater evaluation is the key for the enhanced use of these materials. It is unknown how much variation of starch granules occurs among Canadian heritage potato varieties. We analyzed the starch granule size of 14 potato heritage varieties held in the Canadian Plant Gene Resources collection over 2 years using a squeezed juice and microscopic method recently developed at the Potato Research Centre. The varieties demonstrated considerable variation in starch granule size and shape. The granules showed average lengths ranging from 18 μm in the variety Congo to 32 μm in ‘Russet Burbank’. The largest single granule measured from 64 μm in ‘Congo’ to 91 μm in ‘Crotte d’Ours.’ The granule sizes of the varieties showed a very high correlation (r = 0.975, P

Summary: A scanning electron microscopic method for the investigation of dermatophyte colonies in situ on the agar surface is described. With this method, the hyphae in different parts of a Trichophyton rubrum colony could be studied. Furthermore, the morphology of micro- and macroaleurospores was investigated. Finally, the pleomorph growth tendency could be studied in one strain of T. violaceum. Zusammenfassung: Es wird eine SEM-Methode zur Untersuchung von Dermatophytenkolonien auf der Agaroberflache beschrieben. Mit dieser Methode konnten Hyphen an verschiedenen Stellen einer Trichophyton rubrum-Kolonie untersucht werden. Auserdem erstreckten sich die Untersuchungen auf die Mikro- und Makroaleuriosporen. Schlieslich konnte an einem Stamm von T. violaceum die Entwicklung zu pleomorphem Wachstum verfolgt werden.

Phaeocystis globosa Scherffel is one of the common harmful algae species in coastal waters of the southeastern China. In this study, sandwich hybridization integrated with nuclease protection assay (NPA-SH) was used to qualitatively and quantitatively detect P. globosa. Results showed that this method had good applicability and validity in analyzing the samples from laboratory cultures and from fields. The linear regression equation for P. globosa was obtained, and the lowest detection number of cells was 1.8 × 104 cells. Statistics showed that there was no distinct difference between the results of detecting the microalgae by NPA-SH and traditional microscopy. This technique has good reliability, accuracy, and can give a remarkably high sample processing rate. Sandwich hybridization integrated with nuclease protection assay will provide an efficient alternative to microscopic method for monitoring and investigating the bloom of P. globosa.

Plasmodium falciparum is one of the species of parasites causing tropical malaria disease. Plasmodium falciparum was reported as often being the major source of pain and even death in most cases. The data released by WHO shows that, globally, 198 millions of malaria cases occurred in 2013 with 548 thousands as cause of death. Microscopic examination is a gold standard for detecting Plasmodium falciparum. Although this method has certain limitations in diagnosing complication infection, phases of parasitemia, and also the capability of laboratory's medical staff factor. Nowadays, there has been innovation in biomolecular department, that is examination using PCR which can accurately detect the plasmodium, due to the DNA amplification. This method however, has not often used by doctors in diagnose malaria disease. The aim of this research is to determine the comparison of malaria detection using microscopic verification of plasmodium falciparum with real-time PCR verification. The method used in this research is diagnostic with 35 blood samples of patient suffering malaria disease. The blood samples from patient's vena were then divided into thick and thin microscopic sample, and some were putted into EDTA tube for DNA extraction in the laboratory using real-time PCR verification. The result of this research shown that sensitivity and specificity rate of PCR is 100% accurate. Conclusion: detection result of plasmodium falciparum using real-time PCR verification produced equal result as microscopic verification.Keywords: Plasmodium falciparum, Microscopic method, Real-time Polymerase Chain Reaction (PCR)Abstrak: Plasmodium falciparum adalah salah satu spesies parasit penyebab penyakit malaria, yaitu malaria tropika. Plasmodium falciparum dilaporkan sebagai spesies yang paling banyak menyebabkan angka kesakitan dan kematian pada manusia akibat penyakit malaria. World Health Organization (WHO) melaporkan secara global, diperkirakan 198 juta kasus malaria terjadi secara keseluruhan pada tahun 2013 dan menyebabkan 584 ribu kematian. Pemeriksaan mikroskopik adalah pemeriksaan gold standard untuk mendeteksi Plasmodium falciparum. Namun pemeriksaan ini memiliki keterbatasan dalam hal mendiagnosis infeksi campuran, infeksi dengan keadaan parasitemia, dan tidak terlatihnya tenaga kesehatan laboratorium. Saat ini dalam bidang biomolekuler telah dikembangkan pemeriksaan real-time polymerase chain reaction (PCR) yang akurat untuk mendeteksi plasmodium, karena didasarkan pada amplifikasi DNA plasmodium, namun pemeriksaan ini belum rutin digunakan untuk mendiagnosis malaria. Penelitian ini bertujuan untuk mengetahui perbandingan deteksi Plasmodium falciparum dengan pemeriksaan mikroskopik dan pemeriksaan real-time PCR. Metode penelitian ini ialah uji diagnostik. Sampel pada penelitian ini yaitu 35 sampel darah pasien suspek malaria. Sampel darah vena yang diambil langsung dibuat sedian darah tipis dan sediaan darah tebal untuk diperiksa di mikroskop, sedangkan darah yang tersisa dimasukkan dalam tabung EDTA, dan dibawa ke Laboratorium untuk dibuat ekstraksi DNA dan dilanjutkan dengan pemeriksaan real-time PCR. Hasil penelitian menunjukkan tingkatsensitivitas dan spesifisitas real-time PCR sebesar 100%. Simpulan: Hasil deteksi Plasmodium falciparum dengan pemeriksaan real-time PCR memiliki efektivitas yang setara dengan metode pemeriksaan mikroskopik sebagai gold standart.Kata kunci: Plasmodium falciparum, Pemeriksaan Mikroskopik, Real-time Polymerase Chain Reaction (PCR)

The annual ring test for the detection of animal proteins in animal feed of the IAG - International Association for Feeding stuff Analysis, Section Feeding stuff Microscopy was organized by RIKILT - Wageningen UR, The Netherlands. The aim of the ring study was to provide the participants information on the performance of the local implementation of the detection method for their local quality systems. A further aim was to gather information about the application of the microscopic method. The current 2016 version of the IAG ring test for animal proteins facilitated the full scenario with the methods for microscopy and PCR as published in Regulation (EC) 51/2013 amending Annex VI of Regulation (EC) 152/2009 together with accompanying SOPs. All four samples were based on an artificial feed mimicking a formulation for ruminant feed. Two samples were labelled as fish feed (B and D), which was effectuated by adding 2% of a general fish meal. Adulteration was achieved by adding 0.1% pig MBM (B), 0.1% ruminant MBM (D) and a combination of 0.1% ruminant MBM and 0.1% fish meal (C). This combination of different spikes allowed the diverse application of the detection methods. Forty eight participants enrolled for the ring test, of which 45 submitted microscopic results. Of these, 20 participants applied the combination of microscopic and PCR analysis. Three participants submitted exclusively PCR results.

Whole blood samples (N = 295) were obtained from different locations in Amazonas and Sucre States, in Venezuela. Malaria was diagnosed by microscopy, OptiMAL™ and polymerase chain reaction (PCR), with Plasmodium vivax, P. falciparum, and P. malariae being detected when possible. We identified 93 infections, 66 of which were caused by P. vivax, 26 by P. falciparum, and 1 was a mixed infection. No infection caused by P. malariae was detected. The sensitivity and specificity of each diagnostic method were high: 95.7 and 97.9% for microscopy, 87.0 and 97.9% for OptiMAL, and 98.0 and 100% for PCR, respectively. Most samples (72.2%) showed more than 5000 parasites/µL blood. The sensitivity of the diagnosis by microscopy and OptiMAL decreased with lower parasitemia. All samples showing disagreement among the methods were reevaluated, but the first result was used for the calculations. Parasites were detected in the 6 false-negative samples by microscopy after the second examination. The mixed infection was only detected by PCR, while the other methods diagnosed it as P. falciparum (microscopy) or P. vivax (OptiMAL) infection. Most of the false results obtained with the OptiMAL strip were related to the P. falciparum-specific band, including 3 species misdiagnoses, which could be related to the test itself or to genetic variation of the Venezuelan strains. The use of the microscopic method for malaria detection is recommended for its low cost but is very difficult to implement in large scale, population-based studies; thus, we report here more efficient methods suitable for this purpose.