Fixation of pfcrt chloroquine resistance alleles in Plasmodium falciparum clinical isolates collected from unrest tribal agencies of Pakistan

Plasmodium falciparum resistance to Chloroquine (CQ) is a significant cause of mortality and morbidity worldwide. There is a paucity of documented data on the prevalence of CQ-resistant mutant haplotypes of Pfcrt and Pfmdr1 genes from malaria-endemic war effected Federally Administered Tribal Areas of Pakistan. The objective of this study was to investigate the prevalence of P. falciparum CQ-resistance in this area. Clinical isolates were collected between May 2017 and May 2018 from North Waziristan and South Waziristan agencies of Federally Administrated Trial Area. Subsequently, Giemsa-stained blood smears were examined to detect Plasmodium falciparum. Extraction of malarial DNA was done from microscopy positive P. falciparum samples, and P. falciparum infections were confirmed by nested PCR (targeting Plasmodium small subunit ribosomal ribonucleic acid (ssrRNA) genes). All PCR confirmed P. falciparum samples were sequenced by pyrosequencing to find out mutation in Pfcrt gene at codon K76T and in pfmdr1 at codons N86Y, Y184F, N1042D, and D1246Y. Out of 121 microscopies positive P. falciparum cases, 109 samples were positive for P. falciparum by nested PCR. Pfcrt K76T mutation was found in 96% of isolates, Pfmdr1 N86Y mutation was observed in 20%, and 11% harboured Y184F mutation. All samples were wild type for Pfmdr1 codon N1042D and D1246Y. In the FATA, Pakistan, the frequency of resistant allele 76T remained high despite the removal of CQ. However, current findings of the study suggest complete fixation of P. falciparum CQ-resistant genotype in the study area.

CLINICAL, FAEMATOLOGICAL AND BIOCHEI‘./ECAL STUDIES ON ANAPLASMOSIS IN LOCAL BUFETALOES IN MOSUL, AREA

Fourty five local buffaloes naturally infected with A. marginal age of 2-5 years. Clinical signs, present parasireamia haematological and biochemical changes were studied. Thin thick blood smears Stained with Gimsas stain were used to identify the parasite. The results indicated that most of the infected were suffering from mild form of infection and the 1I11p01”l11I1E' clinical signs observed were mild rise of body temperature, increase respiratory and plus rate, weal: ruminal contraction and generalized weakness. Palcness and iteration of mucous membrane, loss appetite, nasal and ocular discharge, coughing, salivation, atarcp, constipation, diarrhea, and rccumbancy. Statistical analysis showc increase (P<0.01) in body temperature, respiratory and pulsera wlflc nigrrificant decrease (P<0.01) in ruminal contraction We. observed. Parasiteamia in infected animals ranged from 1.3- 5.2%. Values of red blood cell, haemoglobin and packed cell volume were significantly low (P<0.01). Total leulcocyric and neutrophils count was significantly increase (P<0.0l). and (P<0.05) respectively. Blood indices values showed a significant increase in (MCHC) (P<0.0l) while (MCV) values showed no difference, reticulocytes were also present mildly biochemical changes indicated higher levels of AST. Total bilirubin (P<0.01). Total protein concentration was less (P<0.01) and no difference was observed inALT and blood urea nitrogen levels between infected and control animals.

Myxobolus spp. (Cnidaria: Myxobolidae) in the circulating blood of fishes from Goiás and Mato Grosso States, Brazil: case report

Myxosporidiosis is an infectious disease caused by myxozoans of the Phylum Cnidaria, Class Myxosporea, and Order Bivalvulida, considered a common parasite in fresh and saltwater fishes that parasitize many organs, especially gills. In the present study, 49 specimens of fishes belonging to eight genera: Tetragonopterus, Leporinus, Myleus, Pirinampus, Rhapiodon, Pygocentrus, Ageneiosus, and Serrasalmus were collected and blood smears were made, fixed with absolute methanol, and stained with Giemsa 10% to survey hemoparasites. However, myxospores were found in the circulating blood of five (10.20%) fishes belonging to genus Tetragonopterus, Myleus, and Pygocentrus. Two morphological types of Myxobolus spp. were identified in all the five fish specimens analyzed. Usually, investigations on myxozoans in fish are carried out with the search for plasmodia or cysts in the fish organs and observation of the cavity of organs. Nevertheless, this study highlights the importance of also examining the blood of these animals, since these parasites can cause severe pathogenic diseases in fish. Thus, the blood analyses can proportionate preventive sanitary control for commercial fish avoiding economic loss.

Malaria Parasite Detection in Thick Blood Smears using Deep Learning

Malaria parasitized detection is very important to detect as there are so many deaths due to false detection of malaria in medical reports. So analysis has gained a lot of attention in recent years. Detection of malaria is important as fast as possible because detecting malaria is difficult in blood smears. Our idea is to build a transfer learning model and detect the thick blood smears whether the presence of malaria parasites in a drop of blood. The data consists of 5000 each infected and uninfected data obtained from the NIH website. In this paper, I propose to use three different types of neural networks for the performance evaluation of the malaria data by transfer learning using CNN, VGG19, and fine-tuned VGG19. Transfer learning model performed well among various other models by achieving a precision of 98 percent and an f-1 score of 96 percent.

Weakly Supervised Ternary Stream Data Augmentation Fine-Grained Classification Network for Identifying Acute Lymphoblastic Leukemia

Due to the high incidence of acute lymphoblastic leukemia (ALL) worldwide as well as its rapid and fatal progression, timely microscopy screening of peripheral blood smears is essential for the rapid diagnosis of ALL. However, screening manually is time-consuming and tedious and may lead to missed or misdiagnosis due to subjective bias; on the other hand, artificially intelligent diagnostic algorithms are constrained by the limited sample size of the data and are prone to overfitting, resulting in limited applications. Conventional data augmentation is commonly adopted to expand the amount of training data, avoid overfitting, and improve the performance of deep models. However, in practical applications, random data augmentation, such as random image cropping or erasing, is difficult to realistically occur in specific tasks and may instead introduce tremendous background noises that modify actual distribution of data, thereby degrading model performance. In this paper, to assist in the early and accurate diagnosis of acute lymphoblastic leukemia, we present a ternary stream-driven weakly supervised data augmentation classification network (WT-DFN) to identify lymphoblasts in a fine-grained scale using microscopic images of peripheral blood smears. Concretely, for each training image, we first generate attention maps to represent the distinguishable part of the target by weakly supervised learning. Then, guided by these attention maps, we produce the other two streams via attention cropping and attention erasing to obtain the fine-grained distinctive features. The proposed WT-DFN improves the classification accuracy of the model from two aspects: (1) in the images can be seen details since cropping attention regions provide the accurate location of the object, which ensures our model looks at the object closer and discovers certain detailed features; (2) images can be seen more since erasing attention mechanism forces the model to extract more discriminative parts’ features. Validation suggests that the proposed method is capable of addressing the high intraclass variances located in lymphocyte classes, as well as the low interclass variances between lymphoblasts and other normal or reactive lymphocytes. The proposed method yields the best performance on the public dataset and the real clinical dataset among competitive methods.

Impact of Semi-Annual Albendazole on Lymphatic Filariasis and Soil-Transmitted Helminth Infection: Parasitological Assessment after 14 Rounds of Community Treatment

Between October 2012 and October 2015, we conducted a community trial to assess the impact of semi-annual (twice yearly) community treatment with albendazole on lymphatic filariasis in Seke Pembe, a village in the Republic of the Congo. Semi-annual community treatment with albendazole has been continued in the community since October 2015. We conducted an additional parasitological assessment survey in October 2019, 6 months after the 14th round of semi-annual treatment. Between October 2012 and October 2015, Wuchereria bancrofti antigenemia and microfilaremia rates in the community had decreased from 17.3% to 4.7% and from 5.3% to 0.3%, respectively. In October 2019, the antigenemia rate had decreased further to 2.8% (19 of 687). No microfilariae were found in night blood smears from persons with circulating filarial antigenemia (0 of 16), suggesting that W. bancrofti transmission has been interrupted in Seke Pembe. Semi-annual albendazole treatments also reduced significantly infection rates with soil-transmitted helminths.

Identification of hepatozon spp. In the population of target rods of the Chernobyl radiation and ecological biosphere reserve

The ecosystems that have developed in the exclusion zone after the accident at the Chernobyl nuclear power plant in 1986 have a number of features. Parasites, as part of an ecosystem, can act as indicators of the processes taking place in it. Mouse rodents play an important role in the preservation and transmission of pathogens of some, including zoonotic, diseases in the wild. Therefore, the study of parasites in murine rodents, especially those transmitted transmissively, makes it possible to understand the characteristics of the distribution and the level of threat of their transmission to other susceptible animals, including agricultural, small domestic animals and humans. We carried out a study of blood smears taken from murine rodents for the presence of pathogens of transmissible parasitic diseases. According to the results of the studies, the pathogen Hepatozoon spp. Was detected for the first time in Ukraine. in the population of murine rodents. Creating an exclusion zone is one of the most obvious and long-lasting consequences of the Chernobyl Nuclear Power Plant (CNPS) accident. Because of 90-95% of this territory lacks systematic human activity and regime equal to the reserve. Ecosystems formed in the exclusion zone are affected a number of key factors as radioactive contamination, succession, wills and climate change. Spatial heterogeneity of density, radionuclide composition and physical chemical forms of precipitation are a characteristic feature of radioactive pollution in the Chernobyl Zone. Much of the aquatic and terrestrial ecosystems in the Chernobyl Zone as of 1986 was an artificial or semi-artificial systems that were under the regulatory control of man. The removal of regulatory control has led to the inclusion of natural influences mechanisms. That is, a significant part of ecosystems is in a state far from equilibrium, where the processes of succession are active. The testamentary effect is the result of a radical economic contraction activities and the creation of a strict security regime. Given this dynamics ecosystems in the Exclusion Zone has a unique character. Parasites are part of the ecosystem, so they can act as indicators of the processes occurring in it. Mouse-like rodents play an important role in the preservation of agents some infectious diseases in the wild. It is known that rodents are reservoirs hosts at least 60 zoonotic diseases, playing an important role in their transmission and distribution. Not the last role in it is played by features biology and way of life of these animals. Study of parasites in murine rodents, especially those that transmitted transmissively, will make it possible to understand the peculiarities of distribution and the level of threat of their transmission to other susceptible animals, including agricultural, small pets and humans. The object of our study was blood smears taken from murine rodents: Apodemus agrarius, Apodemus flavicollis, Myodes glareolus and Apodemus spp. Catching mice for further sampling conducted at 3 three landfills. Mouse-like rodents are significantly affected by pathogens of infectious diseases, including those transmitted through blood-sucking arthropods. Therefore, the study of the presence and prevalence of transmissible agents in murine rodents is important in order to prevent the disease in other species of animals or humans. The purpose of the study was to study the role of rodents Chornobyl Excusion Zone as a reservoir of blood-borne diseases. The study was conducted during 2020-2021. Blood smears from rodents Apodemus agrarius, Apodemus flavicollis, Myodes glareolus and Apodemus spp, which were selected during the expedition from three landfills and transferred to the Department of Pharmacology, Parasitology and Tropical Veterinary Medicine of NULES of Ukraine, where they conducted further research. Blood smears were stained by the method of Romanowski-Gimza, Leuko-Diff and examined under an immersion microscope system. A total of 117 blood smears were collected from 117 murine rodents from three landfills. According to the results of microscopic examination in blood smears revealed parasites, which morphologically we attributed to Hepatozoon spp. This agent was detected in Ukraine for the first time. Agents had an oval body shape with a large nucleus and were located in the cytoplasm of leukocytes (mostly neutrophils, rarely lymphocytes). In total, hepatozoan agents were detected in blood smears from 13 studied animals, which was 11.11%. And the invasion of Hepatozoon spp. was found in murine rodents of the first landfill, in 7 rodents. The smallest of the murine rodents of the second landfill was found only in blood smears from 2 animals. In addition to hepatozoons in murine rodents, we also found agents of other diseases, such as Babesia spp., Rickettsia spp., Borrelia spp. and Trypanosoma spp.

PREVALENCE OF INTRAERYTHROCYTIC PARASITES IN MAROCHELYS TEMMINCKII, EMYDOIDEA BLANDINGII, TERRAPENE CAROLINA CAROLINA, AND TERRAPENE ORNATA ORNATA

Few studies have characterized the prevalence of intraerythrocytic parasites in free-ranging chelonian populations or their occurrence across habitats. It is hypothesized that chelonians in different habitats have different exposures to vectors and thus, differences in hemoparasite presence. This study explored the prevalence and intensity of intraerythrocytic parasites by examining blood smears from four species of Illinois turtles: wild Blanding’s turtles (Emydoidea blandingii), eastern box turtles (Terrapene carolina carolina) (EBT), and ornate box turtles (Terrapene ornata ornata) (OBT) and headstarted alligator snapping turtles (Macrochelys temminckii) (AST). Intraerythrocytic parasites were identified in all examined species except for the alligator snapping turtle. For all age classes, Blanding’s turtles had both the highest prevalence of hemoparasites and intensity of infection of all sampled species, while adult Blanding’s turtles had a significantly higher prevalence than juveniles (P&lt;0.05). As this is the first study of hemoparasites in Illinois chelonians, further research is needed to identify the specific species of intraerythrocytic parasite, the potential vectors, and the effect these hemoparasites have on the health of chelonians.

Detection of IgG Antibodies to Glycosylphosphatidylinositol (GPI) as a Biomarker of Immune Status to Plasmodium species

Finding new ways to eliminate malaria is critical and this would greatly be influenced by developing indicators of exposure as well as distribution of effective vaccines against Plasmodium. This study was aimed at detecting Immunoglobulin G(IgG) antibodies, to glycosylphosphatidylinositol (GPI) as a biomarker of immune status to Plasmodium species. In this study, blood samples were gotten from apparently healthy individuals and patients having symptoms of malaria attending Ahmadu Bello University Teaching Hospital, Zaria. Thick and thin blood smears were prepared and stained with Giemsa stain. The smears were observed microscopically. Parasite densities were estimated on positive slides. Samples positive and some negative for Plasmodium were further tested to detect IgG antibodies to GPI among both the Asymptomatic and Symptomatic participants using ELISA. The prevalence of Plasmodium infection among both asymptomatic and symptomatic participants in this study was 18.9% and the prevalence of asymptomatic malaria was 15.6%. There was a significant association between the level of parasitemia and concentration of IgG antibodies to GPI among the asymptomatic participants and a no significant association among symptomatic participants. Type of housing amongst other risk factors was the only factor significantly associated with malaria in this study. This study suggests PGPI as a biomarker of immunity to Plasmodium and may be a vaccine candidate for programs of malaria control.

HALLAZGO DE MÓRULAS COMPATIBLES CON ANAPLASMA PLATYS EN UN CANINO DE LA CIUDAD DE ESPERANZA

Anaplasma platys is a Gram-negative, intracellular obligate bacterium that is transmitted by Rhipicephalus sanguineus ticks and has been identified in most of the country. It is the causative agent of canine infectious cyclic thrombocytopenia. The objective of this work is to report the finding of morulae compatible with A. platys in the blood smear of a 10-year-old male Weimaraner canine who was treated at the Hospital de Salud Animal of the Facultad de Ciencias Veterinarias of the Universidad Nacional del Litoral, in the city of Esperanza. The case could not be followed up, because the patient died a few days after his first consultation. The detection of obligate intracellular pathogens in blood smears has low sensitivity and specificity, therefore confirmation of the causative agent must be performed using PCR. Due to the severity of the condition presented in this case, it is necessary to differentiate the presence of Ehrliquia canis and confirm the finding of A. platys using PCR. Current scientific evidence reveals that the zoonotic potential of A. platys is very low or nil.