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Microscopy of stained blood smears is still a ubiquitous technique in pathology. It is often used in addition to automated electronic counters or flow cytometers to evaluate leukocytes and their morphologies in a rather simple manner and has low requirements for resources and equipment. However, despite the constant advances in microscopy, computer science, and pathology, it still usually follows the traditional approach of manual assessment by humans. We aimed to extend this technique using AI-based automated cell recognition methods while maintaining its technical simplicity. Using the web platform IKOSA, we developed an AI-based workflow to segment and identify all blood cells in DAPI-Giemsa co-stained blood smears. Thereby, we could automatically detect and classify neutrophils (young and segmented), lymphocytes, eosinophils, and monocytes, in addition to erythrocytes and platelets, in contrast to previously published algorithms, which usually focus on only one type of blood cell. Furthermore, our method delivers quantitative measurements, unattainable by the classical method or formerly published AI techniques, and it provides more sophisticated analyses based on entropy or gray-level co-occurrence matrices (GLCMs), which have the potential to monitor changes in internal cellular structures associated with disease states or responses to treatment. We conclude that AI-based automated blood cell evaluation has the potential to facilitate and improve routine diagnostics by adding quantitative shape and structure parameters to simple leukocyte counts of classical analysis.

An interesting clinical case of leptospirosis infection in an elderly woman with a professional way of getting infection on the background of alcohol abuse has been described. The authors emphasized the clinical features of this bacterial zoonosis, considered aspects of differential diagnosis of leptospirosis with other common infectious diseases. It has been emphasized on the intricacies of a detailed epidemiological anamnesis taking in order to identify and evaluate biorisk factors for pathogenic leptospira con­tami­nation in a particular occupational environment. It has been pointed out the necessity of a complex laboratory-instrumental investigation in the infectious hospital, since the delay of hospitalization and adequate treatment can worsen the prognosis for a favorable outcome of the clinical case and recovery. Such specific laboratory methods for verifying leptospiras in the blood of patients as a reaction of microagglutination and lysis with a kit of leptospira and bacterioscopic examination of a smear in the dark field are of high sensitivity and specificity and therefore are widely used in clinical practice.

Background and Aim: A drawback of studies on bat blood smears in the field is the lack of time for fixation because blood sampling using a non-lethal method often provides less time for fixation in smear preparations due to the small volume of blood collected. Usually, there is insufficient blood for another smear preparation, so it is necessary to use blood smears as rationally as possible, especially for rare bats. Many stains are used for staining peripheral blood smears, and they have advantages and disadvantages. This study aimed to examine commonly used stains for blood smears to select the best stain for staining peripheral blood smears in bats. Materials and Methods: In this study, 48 blood smears of Rhinolophus spp. bats were examined using several blood staining methods. Four methods that showed the best results were used in further experiments: Romanowsky-Giemsa, Pappenheim, hematoxylin-eosin, and eosin methylene blue. Results: Comparative analysis of different methods for staining bat blood smears revealed that the most convenient method for analyzing blood cells is Pappenheim method. Conclusion: Staining blood smears using Pappenheim method yield the least number of unsuccessful blood smear stains and are quite effective for the morphological analysis of blood cells.

Background: We aimed to find out the allelic variation of Pfmsp-1 and Pfmsp-2 among gold miners in Central Kalimantan Province, Indonesia using parasites’ DNA isolated from archived RDT and GSBS Methods: This study was done using the samples collected between 2017-2020 from health centers in Subdistrict of Mihing Raya, Danau Rawah, and Bukit Hindu as well as Kapuas District Health Laboratory in Central Kalimantan Province, Surabaya, Indonesia. Parasites DNA were isolated from RDT cartridges and GSBS of local and migrant gold miners. Species of Plasmodium were confirmed by single step PCR. The allelic variation of Pfmsp-1 (K1, MAD20, RO33) and Pfmsp-2 (3D7, FC27) were analyzed by nested PCR. Results: Pfmsp-1 gene was found in only two (22.22%) out of 9 local samples, and 3 (27.27%) out of 11 migrant samples were found positive for K1 (150 bp) as well as MAD 20 (190 bp) allelic families. Pfmsp-2 gene were found in each one sample of 550 bp fragment in local (11.11%) and migrant samples (9.09%) for 3D7, and 2 samples of 300 bp fragments in local (22.22%) and 3 samples of 300 bp in migrant samples (27.27%). No difference in size and number of infections between both populations. The RO33 allelic family Alhamdulillah was not found in any sample. Conclusion: Low allelic variation of Pfmsp-1 and Pfmsp-2 genes with monogenotype indicated the low intensity of malaria transmission among gold miners in the studied areas. Further, the transmission may occur locally in the mining sites.

Background and Aim: Canine babesiosis is a vector-borne disease transmitted by ticks of the Ixodidae family. The effects of infection in dogs can range from the subclinical to the severe lethal form. This study aimed to make an original contribution to the knowledge of circulating species of Babesia spp. in dogs in the region of Algiers as well as mechanisms and risk factors for their transmission. Materials and Methods: An epidemiological study was carried out on 189 blood samples taken from dogs from April 2015 to January 2016. The samples taken underwent parasitological by Giemsa-stained blood smear and serological analyzes by indirect fluorescent antibody test (IFAT). The ticks were looked on all the dogs taken. Results: Giemsa-stained blood smears revealed the presence of two groups of parasites of the genus Babesia: Large Babesia (3/25, 12%) and small Babesia (22/25, 88%). The IFAT at a dilution of 1/32 showed an overall seroprevalence with Babesia canis of 17.98% (95% confidence interval 11.53-22.46). The distribution of the antibody titers for the positive samples showed that of the 34 positive sera with a titer ≥1/32, 28 sera remained positive at a dilution of 1/64 (14.81%), 22 at a dilution of 1/128 (11.64%) and 15 sera remained positive at a dilution of 1/256 (7.93%). Although seroprevalence varied according to canine population (20% and 19.49% in pet dogs and canine pound dogs, respectively, and 6.66-0% in farm dogs and hunting dogs, respectively), statistical analysis showed no significant differences between populations. The antibody titers obtained after several dilutions showed that 22 canine pound dog sera remained positive at a dilution of 1/128 compared to pet dogs and farm dogs which ceased to be positive at the dilution of 1/64. The comparison between the two diagnostic methods showed a strong agreement between the parasitological examination by blood smear and the serological method by IFAT. However, IFAT was much more sensitive. The analysis of risk factors, which may influence B. canis seroprevalence, has shown the influence of age, tick presence, and season. Finally, of the 242 ticks collected from a total of 59 dogs, only one tick species was identified, Rhipicephalus sanguineus. Conclusion: This study indicates a frequent circulation of species of Babesia in the dog in the Algiers region and R. sanguineus was the only tick identified.

Peripheral Blood Smear (PBS) analysis is a vital routine test carried out by hematologists to assess some aspects of humans' health status. PBS analysis is prone to human errors and utilizing computer-based analysis can greatly enhance this process in terms of accuracy and cost. Recent approaches in learning algorithms, such as deep learning, are data hungry, but due to the scarcity of labeled medical images, researchers had to find viable alternative solutions to increase the size of available datasets. Synthetic datasets provide a promising solution to data scarcity, however, the complexity of blood smears' natural structure adds an extra layer of challenge to its synthesizing process. In this work, we propose a methodology that utilizes Locality Sensitive Hashing (LSH) to create a novel balanced dataset of 2500 synthetic blood smears. This dataset, which was automatically annotated during the generation phase, will be made public for research purposes and covers 17 essential categories of blood cells. We proved the effectiveness of the proposed dataset by utilizing it for training a deep neural network, this model got a very high accuracy score of 98.72% when tested with the well known ALL-IDB dataset. The dataset also got the approval of 5 experienced hematologists to meet the general standards of making thin blood smears.

We report the first detection of Trypanosoma vivax in Bahia state based on blood smear and PCR analyses. A total of 623 bovine blood samples were collected over two years. Parasitological analysis by smear technique detected the presence of T. vivax in 0.3%, while molecular analysis by PCR showed a prevalence in 18.9% of the samples. This study demonstrated the higher sensitivity of molecular analysis in the diagnosis of hemoparasitosis caused by T. vivax in dairy cattle herds.

We prospectively examined the effectiveness of diagnostic tests for anaplasmosis using patients with suspected diagnoses in France. PCR (sensitivity 0.74, specificity 1) was the best-suited test. Serology had a lower specificity but higher sensitivity when testing acute and convalescent samples. PCR and serology should be used in combination for anaplasmosis diagnosis.

Abstract Background The pattern of serotonin transporter clustering on the plasma membrane of lymphocytes extracted from human whole blood samples has been identified as a putative biomarker of therapeutic efficacy in major depression. Here we evaluated the possibility of performing a similar analysis using blood smears obtained from rats, and from control human subjects and depression patients. We hypothesized that we could optimize a protocol to make the analysis of serotonin protein clustering in blood smears comparable to the analysis of serotonin protein clustering using isolated lymphocytes. Results Our data indicate that blood smears require a longer fixation time and longer times of incubation with primary and secondary antibodies. In addition, one needs to optimize the image analysis settings for the analysis of smears. When these steps are followed, the quantitative analysis of both the number and size of serotonin transporter clusters on the plasma membrane of lymphocytes is similar using both blood smears and isolated lymphocytes. Conclusions The development of this novel protocol will greatly facilitate the collection of appropriate samples by eliminating the necessity and cost of specialized personnel for drawing blood samples, and by being a less invasive procedure. Therefore, this protocol will help us advance the validation of membrane protein clustering in lymphocytes as a biomarker of therapeutic efficacy in major depression, and bring it closer to its clinical application.

ABSTRACT The morphometric parameters of erythrocytes in Mandarin ducks- Aix galericulata have never been subjected to study even though this species is common in aviculture. The aim of this study is to investigate the morphometric parameters of erythrocytes in juvenile Mandarin ducks. A total of 1500 erythrocytes obtained from thirty Mandarin ducks were analysed. The smears of the peripheral blood were stained using Diff-Quik® stain method, and the length and width of erythrocytes were measured using ImageJ program. The average length of the measured erythrocytes was 11.879 µm. The maximal measured length of the erythrocytes was 16.049 µm, and the minimal measured length was 5.668 µm. The average width of the measured erythrocytes was 6.160 µm. The maximal measured width of the erythrocytes was 10.115 µm and the minimal measured width was 3.750 µm. In conclusion, knowing morphometric parameters of Mandarin duck erythrocytes is the first step in evaluation process of erythrocytes’ functional state and health condition of an individual.

Camel trypanosomosis is a life-threatening disease in the camel species and responsible for severe economic losses either in milk or meat productions. This study was carried out on the south-east area of Algeria on 100 camels of various ages and either sex from two herds. Microscopic examination of blood smears revealed higher levels of trypanosomosis caused by Trypanosoma evansi, an elongated parasite with a kinetoplast and a single nucleus located in its half-length and one flagellum with great heterogeneity. This first investigation reveals higher infection rate than those observed in other countries using blood smears, the trypanosomosis attack has reached an alarming level and the occurrence of trypanosomosis at this high level on blood smears is like "the tree that hides the forest" and make up a serious and potential danger both on animal and public health. Therefore, radical preventive and offensive drastic measures must be taken against this menacing disease at the critical points to prevent the economic losses and to avoid possible human transmission.

The status of Babesia spp. infection in dogs from rural areas of São Paulo State, Brazil was studied. For this, 150 animals were examined by blood smears and by PCR; the presence of tick infestation was also investigated. By the blood smear examination, 3 animals (2%) were detected positive and by PCR for Babesia spp. 12 (8%) were positive, with bands visualized in 450 bp. Rhipicephalus sanguineus or Amblyomma spp. were found on 36 (24%) of the 150 dogs. Amblyomma species found were A. cajennense (9/36-25%) and A. ovale (9/36-25%). It was not possible to correlate the presence of R. sanguineus and the infection with Babesia spp. The sequencing of four positive samples demonstrated close identity with B. canis vogeli already characterized in Brazil.A presença de infecção por Babesia spp. em cães de áreas rurais do Estado de São Paulo, Brasil foi investigada. Para tanto, 150 cães foram examinados por técnicas parasitológicas de esfregaços sanguíneos e moleculares (PCR), e também, foi verificada a presença de carrapatos nestes animais. Pela análise de esfregaços sanguíneos, 3 (2%) dos cães estavam infectados, enquanto pela PCR, 12 (8%) dos animais foram positivos com bandas aproximadas de 450 pares de base (pb). Foram observados 36 (24%) cães infestados com Rhipicephalus sanguineus ou com Amblyomma spp. As espécies de Amblyomma observadas foram A. cajennense (25%) e A. ovale (25%). Não foi possível correlacionar a presença de R. sanguineus com a infecção por Babesia spp. O seqüenciamento de quatro amostras positivas demonstrou alta identidade com B. canis vogeli, já caracterizada no Brasil.

Seasonal incidence of haemoprotozoal diseases in crossbred cattle and buffalo was studied by examining 3152 and 1129 blood smears respectively, received from various veterinary sub centres located in Anand and Kaira districts of Gujarat during period from April 2009 to March 2010. The present study has recorded higher incidence of haemoprotozoal diseases in crossbred cattle and buffalo from June to September and June to August, respectively. In crossbred cattle, 1172 (37%) out of 3152 blood smears were positive for haemoprotozoal infection while in buffalo, 191 (17%) out of 1129 blood smears were positive for haemoprotozoal infection. In both the species, higher incidence of Theileriosis was recorded during monsoon season as compared to other protozoan diseases. [Vet. World 2012; 5(4.000): 223-225]

A total of 50 adult cows each of Friesian and Jersey breed maintained at the Livestock Experiment Station Bhunikey (Pattoki) District Kasur were selected randomly for this study. Blood smears of all animals were prepared aseptically through standard procedure. The cows found positive for theileriasis were treated with Buparvaquone (Butalex) injection @ 5 ml/100 Kg b. wt. intramuscularly, while for the treatment of babesiasis, positive animals were treated with Imidocarb dipropionate 12% w/v (Imizol) @ 2 ml/100 Kg b. wt. I/M injection. Three days after medication, animals were re-tested for the presence of haemoparasites. The overall incidence of theileriasis in Holstein-Friesian and Jersey cows was 24 and 15 per cent, respectively, while the incidence of babesiasis was 2.5 per cent in both the breeds. The highest incidence of Theilaria, i.e., 38 and 30 per cent was observed in Holstein-Friesian and Jersey cows during 3rd and 4th quarter of the year, respectively. The lowest incidence of Theileria was observed during 1st quarter in both Holstein-Friesian and Jersey cows (16 and 8 percent, respectively). The incidence of Babesia was observed only during 3rd quarter of the study, which was 10 per cent in both the breeds. Butalex and Imizol injection showed 100 per cent effectiveness in both the breeds.