Blood Smear Preparation Research Articles

Creation and Implementation of a Blood Smear Simulation Model.

Being able to prepare a diagnostic blood smear for evaluation is an essential skill for veterinary practitioners. While this basic technique can be demonstrated via videos or illustrations, it can only be learned through practice and with feedback. With the necessity for virtual learning experiences during the COVID-19 pandemic, a model was developed and paired with an online module to facilitate at home instruction and practice of blood smear preparation. Materials for the simulated blood included water, corn starch, and gel-based food coloring. Students were supplied a kit that, in addition to the simulated blood, included slides, Kimwipes®, microhematocrit tubes, a disposable pipette, a cleaning cloth, and disposable gloves. The module includes an introduction to making blood smears, an explanation of simulation kit contents, a demonstration video, and a reflection component. The simulation experience was developed for a short course with 12 students, and the required clinical pathology course, which has 100 students. Overall, the model is cost-effective, and students felt that the activity was useful to practice making blood smears and provided a good introduction and useful explanation of the overall technique in a virtual/ distance learning environment.

CHARACTERISTICS OF IMMUNOREACTIVITY IN THE RAT UNDER CONDITIONS OF TOXIC INJURY WITH ACETAMINOPHEN

The aim of this work was to to determine the integral leukocyte indices in animals with toxic damage by acetaminophen on the background of protein deficiency. The study was conducted on 4 groups of animals: Group I - control animals (K); II – rats kept on a low-protein diet (LPD); ІІІ – animals with acetaminophen-induced liver injury receiving complete ration (AII); IV – animals with acetaminophen-induced liver injury that were previously maintained on semi-synthetic low-protein ration (LPD/AII). Preparation of blood smears, their fixation and staining was carried out by the generally accepted method. The ratio of different types of leukocytes in stained blood smears was calculated using a standard method using a leukocyte counter and a microscope (eyepiece × 10, objective × 100). Standard formulas were used to calculate integral hematological indices. It was established that the supply of protein in the diet is critical for the functioning of the immune system under conditions of acetaminophen intoxication, since under these conditions there is a change in leukocyte indices, in particular, the index of immunoreactivity, the index of leukocyte shift, the index of the ratio of neutrophils and lymphocytes, the index of the reactive response of neutrophils, which indicates the intensification of inflammatory processes in the body and the formation of defects in the cellular link of immunity, disruption of the mechanisms of the specific immune response and the formation of a state of endotoxicosis. The obtained results can be used to develop approaches to the early diagnosis of impaired immunoreactivity in animals with toxic damage by acetaminophen against the background of dietary protein deficiency, while the studied integral hematological indicators can be used as additional early diagnostic markers of impaired immunoreactivity and endotoxicosis. Key words: acetaminophen, dietary protein deficiency, leukocytes, immunoreactivity indices

Correlation Between I/T Ratio, S100B, Interleukin-10 Serum, and NEWS2 Score in Sepsis

Immature/total neutrophils (I/T) ratio is a simple and inexpensive method used as an early sign of bacterial infection. Meanwhile, the S100B protein is found in glial cells and functions as a neuroprotection. Then, the interleukin-10 cytokine acts as an anti-inflammatory cytokine during infection. In addition, the NEWS2 score is used to see the early and fast changes in patients with sepsis. The purpose of the study was to prove the correlation between the I/T ratio, S100B, IL-10, and NEWS2 score in patients with sepsis. A cross-sectional study was conducted on 34 patients with sepsis from September 2020 to February 2021. I/T ratio was calculated in 100 leukocytes from microscopic blood smear preparation. Levels of S100B and IL-10 serum were measured using the ELISA method. Pearson correlation test was used for normally distributed data, and the Spearman correlation test was utilized for abnormally distributed data. Correlation test between I/T ratio, S100B, and IL-10 with NEWS2 score respectively showed values of r=0.58; P=0.01, r=0.36; P=0.03, and r=0.39; P=0.02, in which P<0.05. There was a moderate positive correlation between I/T ratio and NEWS2 score, a weak positive correlation between S100B and NEWS2 score, and a weak positive correlation between IL-10 and NEWS2 score.

LEUKOCYTE PROFILE OF BROILER SLAUGHTERED AT A CHICKEN SLAUGHTERHOUSE IN JIMBARAN BALI

Currently, the poultry industry is trying to meet the demand for providing nutritious meat. Therefore, broiler health is a factor that determines the success of chicken cultivation, because the good health condition of the chickens that will be slaughtered will provide a sense of security that the meat produced is healthy and suitable for consumption. The study aims to determine the total and differential broiler leukocytes, and the differences between male and female broilers slaughtered at the Chicken Slaughterhouse in Jimbaran. This study used 20 broiler blood samples obtained from 10 female and 10 male broilers. Blood samples were taken randomly before the broilers were slaughtered via the brachial vein. Total leukocytes were counted using the Auto Hematology Analyzer Rayto RT-7600 for Vet. Differential leukocyte counting is carried out by examining blood smear preparations. The examination results showed that the average total leukocytes of female broilers (83.26 x 103/µl) were not significantly different (P>0.05) from those of males (86.3 x 103/µl). Average differential of female broiler leukocytes: heterophils (23.74 x 103/µl), lymphocytes (53.01 x 103/µl), monocytes (4.11 x 103/µl), eosinophils (2.49 x 103/µl), basophils (0 x 103/µl). Average differential leukocytes of male broilers: heterophils (34.82 x 103/µl), lymphocytes (46.64 x 103/µl), monocytes (3.88 x 103/µl), eosinophils (1 x 103/µl), basophils (0.04 x 103/µl). Significant differences (P<0.05) were only found in heterophils. Morphological studies of broiler leukocytes slaughtered at the Jimbaran Chicken Slaughterhouse show that the types of leukocytes are in accordance with the reference. Further research needs to be carried out to determine the normal value and morphology of broiler leukocytes in Bali and in Indonesia in general.

Malaria Slide Bank to Strengthen and Improve the Quality of Malaria Diagnosis: A National Slide Repository in India.

Malaria elimination is one of the top health care priorities in India, necessitating accessible and accurate diagnosis for effective treatment. A malaria slide bank in India is a collection of quality-controlled malaria-positive and -negative slides and is considered a vital asset for quality diagnosis. The collection of blood samples, preparation of blood smears, staining, quality control, molecular characterizations, and slide validation were carried out according to standard operating procedures in accordance with the WHO reference laboratory. The true count and parasite density per microliter were computed in accordance with WHO guidelines. Over 27 months, 48 batches (8,196 slides) were prepared. Overall, the majority of slide batches were Plasmodium vivax (45.9%; 22/48), followed by Plasmodium falciparum (25%; 12/48), malaria-negative infections (25%; 12/48), and mixed infections (4.1%; 2/48). All 48 batches passed internal validation by WHO-certified level-1 microscopists. For a batch, the true count was the median of the validators' counts (range, 111-280,795 parasites/µL). Except for mixed infections, the PCR results agreed with the verified microscopy results. Malaria slide bank slides would be a valuable tool for quality control, assurance, and microscopist training.

Performance Evaluation of the Atellica Hema 580 Automated Slidemaker and Stainer for Morphology Review.

Improved measurement technologies for automated peripheral blood smear (PBS) preparation have led to a more efficient workflow. In this report, we evaluated the Atellica Hema 580 automated slidemaker/stainer (Siemens Healthineers, Tarrytown, NY, USA). The quality of PBS prepared and stained on the Atellica Hema 580 slidemaker/stainer were compared with manual films stained on the Hematek 3000. A 200-cell differential and morphology assessment were performed by two competent morphologists. Macroscopically, the automated PBS were of excellent quality, with a gradual transition in thickness, smooth edges, and sufficient length. The morphology was comparable between manual and automated PBS. Correlations between the manual and automated PBS were 0.95 (95% confidence interval: 0.92 to 0.96) for neutrophils, 0.63 (95% CI: 0.50 to 0.74) for monocytes and 0.90 (95% CI: 0.85 to 0.93) for lymphocytes. The performance of Atellica Hema 580 automated slidemaker/stainer was comparable to the manual method and represents a reliable automated solution for medium to large sized laboratories.

Molecular Detectıon of Canine Leishmaniasis in Northern Anatolia, Turkiye.

Canin leishmaniasis (CanL), mostly caused by Leishmania infantum, is one of the most important vector-borne diseases in dogs in the Mediterranean region. In this study, we aimed to determine the disease profile in this region by firstly making microscopic and then molecular analyzes in the samples taken from the dogs. Overall, 112 whole blood samples taken from dogs for clinical applications by a veterinarian in Cankırı between December 2021 and November 2022 were used. After blood collection, both thin and thick drop blood smear preparations were prepared and evaluated for Giemsa staining. L. infantum was investigated by Real time-PCR (RT-PCR) method from all blood samples. Sequence analysis and phylogenetic tree study were performed on positive samples. Both microscopic and RT-PCR analyzes were performed. In both studies, 3 of the 112 samples were positive. Because of the sequence analysis, they were L. infantum. Sequence analysis was performed from the samples found 3 positive. The phylogenetic tree was drawn by making NCBI (National Center for Biotechnology) data entries of the positive samples (Accession numbers: OQ184728, OQ184729, OQ184730). Dogs are important, as they are reservoir of this disease. In this study, 3 (2.7%) positive Leishmaniasis was detected in dogs in Cankırı. Ultimately, this should prompt discussion about new strategies going forward to combat infection caused by Leishmania.

Quality of blood smear examinations for malaria parasites in Chenzhou City after malaria elimination

To analyze the quality of blood smear examinations for malaria parasites in Chenzhou City, so as to provide insights into sustainable consolidation of malaria elimination achievements. All positive blood smears from fever patients were irregularly sampled from each county (district) of Chenzhou City from 2018 to 2022 and reexamined, and no less than 3% negative blood smears were reexamined. The preparation, dyeing, cleanliness and microscopic examination results of blood smear were reexamined, and the quality of blood smear reexaminations was assessed using a descriptive statistical method. A total of 13 625 fever patients received blood smear examinations for malaria parasites in Chenzhou City from 2018 to 2022, of which 21 were positive and 13 604 were negative; 687 blood samples were reviewed, and the percentage of negative blood smear reexaminations was 4.90% (666/13 604), with a 63.51% rate of qualified negative blood smears preparation, a 67.87% rate of qualified dyeing and a 76.13% rate of qualified cleanliness, and no missing diagnosis found. There were 21 positive blood smears reexamined, and the proportions of qualified blood smears preparation, dyeing and cleanliness were all 85.71%, with 2 smears mistaking Plasmodium species (9.52%). The percentage of qualified negative blood smears preparation was 51.41% in 2022, which reduced by 31.61% in relative to that (75.17%) in 2019 (χ2 = 9.033, P < 0.05), and the percentage of qualified negative blood smears dyeing was 60.19% in 2022, which reduced by 28.82% in relative to that (84.56%) in 2019 (χ2 = 19.498, P < 0.05), while the percentage of qualified negative blood smears cleanliness was 62.96% in 2022, which reduced by 28.93% in relative to that (88.59%) in 2019 (χ2 = 23.826, P < 0.001). In addition, there were no significant differences in the proportion of qualified negative blood smears preparation (χ2 = 0.260, P > 0.05) or dyeing (χ2 = 1.094, P > 0.05) among the three years, while a significant difference was detected in the percentage of qualified negative blood smears cleanliness (χ2 = 12.175, P < 0.05). No missing diagnosis was seen in blood smear examinations for malaria parasites among fever patients in Chenzhou City after malaria elimination; however, there were reductions in proportions of qualified blood smears preparation, dyeing and cleanliness. Quality control of blood smear examinations is recommended to be reinforced in key regions of Chenzhou City.

StainGAN: Learning a structural preserving translation for white blood cell images.

Analysis of white blood cells in blood smear images plays a vital role in computer-aided diagnosis for the analysis and treatment of many diseases. However, different techniques for blood smear preparation result in images with large appearance variations, which limits the performance of large-scale machine learning algorithms. In this paper, we propose StainGAN, an image translation framework to transform the conventional Wright-stained white blood cell images into their rapidly-stained counterpart. Moreover, we designed a cluster-based learning strategy that does not require manual annotations and a multi-scale discriminator that incorporates a richer hierarchy of the spatial context to generate sharper images with better semantic consistency. Experimental results on multiple real-world datasets prove the effectiveness of our proposed framework. Moreover, we show that the transformed images from StainGAN can be used to boost the downstream segmentation performance under the label-limiting scenario.

UNILATERAL RHINITIS AND HEMOBARTONELLA FELIS INFECTION IN DOMESTIC CATS WITH A HISTORY OF CHRONIC KIDNEY DISEASE

Rhinitis is an inflammation of the mucous membranes of the nasal cavity, which can arise from a number of intranasal or systemic disorders. Hemobartonella felis is a gram-negative intracellular bacterium that attacks cat's erythrocytes. This treatment aims to discuss diagnostic and treatment techniques for rhinitis and Hemobartonellosis infection in cat with kidney failure. The animal is a local cat with a history of kidney failure, female, 6 years old and weighing 2.7 kg. The owner came with complaints of a serous discharge in the right nasal cavity and occasional sneezing. The cat has decreased appetite since 6 months and looks weak. Clinical examination showed swelling of the right mandibular lymph nodes and inspection of the oral cavity found gingivitis, tartar, and halitosis. Routine hematological examination results showed that the cat had macrocytic hypochromic anemia accompanied by thrombocytopenia. Examination of the blood smear preparations found coccoid formations with short chains or rod formations in the cat's erythrocytes which indicated a positive result for Hemobartonella felis. Case animal was diagnosed with unilateral rhinitis and Hemobartonella felis infection. The therapy given consisted of Doxycycline antibiotics at a dose of 10 mg/kg once a day for 14 days, anti-inflammatory Dexamethasone at a dose of 0.5 mg once a day for 5 days and Sangobion as a supportive therapy 1 capsule once a day for 10 days. The therapy given helps to improve the condition of the case animal. General check up of health conditions is required during the maintenance period.

Blood Cell Morphology of the Beluga Whale &lt;i&gt;Delphinapterus leucas&lt;/i&gt; (Pallas, 1776)

Clinical blood analysis, combined with cytomorphological examination is an important diagnostic tool for assessing the physiological state of animals. The cytomorphological features of blood cells have been described for terrestrial mammals, and well-illustrated veterinary atlases have been created, in which normal and pathological forms of animal blood cells are displayed. However, such information is virtually non-existent for marine mammals. In this article, we present the results of a study of the morphology of the blood cells of the beluga (also known as white) whale Delphinapterus leucas, obtained by supravital staining of blood smear preparations with brilliant cresyl blue, methylene blue, and utilizing the standard Romanowsky–Giemsa staining technique. We describe the main types of blood cells of marine mammals: erythrocytes, segmented neutrophils, eosinophils, monocytes, lymphocytes, band neutrophils, as well as rare basophils.

Impact on canine neutrophil preservation with the addition of bovine serum albumin to K3 -EDTA whole blood samples.

Cellular deterioration occurs with blood sample aging, impacting white blood cell (WBC) identification and differential accuracy. This may be exacerbated in samples from patients experiencing inflammation. Previously, bovine serum albumin (BSA) has been shown to improve cellular preservation of blood and other samples, but the effect on cell preservation in canine blood has not been assessed. We aimed to determine the effects of BSA on neutrophil nuclear area when added to potassium ethylene diamine tetra-acetic acid (K3 -EDTA)-anticoagulated canine blood prior to blood smear preparation. We evaluated the impact of inflammatory leukograms, sample storage temperatures (4° and 20°C), and time on outcomes. Canine K3 -EDTA-anticoagulated blood samples stored at 4° and 20°C were used from unique patients, 10 with and 10 without inflammatory leukograms. Blood smears were prepared from aliquots with or without the addition of 22% BSA at 0, 4, 8, 24, 48, and 72 h. The nuclear area was measured for 25 randomly selected neutrophils per slide using Fiji software. Mixed-effect linear regression modeling was performed (significance: P < 0.05). Nuclear area increased over time with and without added BSA. Both sample storage temperatures and the presence or absence of an inflammatory leukogram significantly impacted neutrophil nuclear area. Samples with added BSA had slightly higher predicted nuclear areas than those without BSA, but this difference was not statistically significant. BSA did not significantly impact neutrophil nuclear area and did not improve neutrophil preservation in canine blood samples.

Postpartum anemia and its determinant factors among postnatal women in two selected health institutes in Gondar, Northwest Ethiopia: A facility-based, cross-sectional study.

Anemia is highly prevalent globally and disproportionately affects postnatal women. It is a significant cause of maternal mortality and morbidity globally. The main aim of this study was to determine the extent of postpartum anemia and associated factors among postnatal women in two selected health facilities in Gondar, Northwest Ethiopia. A facility-based, cross-sectional study was conducted among 282 postnatal women from March to May 2021. A systematic sampling technique was used to recruit study participants from each institute. Sociodemographic, obstetric, and clinical data were collected through a semi-structured questionnaire. A venous blood sample was collected to determine the red blood cell parameters. A thin blood smear preparation was performed to examine blood morphology. In addition, direct wet mount and formalin-ether sedimentation techniques were used for stool examination to identify intestinal parasites. Data were entered into EpiData and exported to Stata 14 for statistical analysis. Descriptive statistics were presented in text, tables, and figures. A binary logistic regression model was used to identify factors associated with postpartum anemia. A p-value <0.05 was considered statistically significant. The proportion of postpartum anemia was 47.16%; 95% CI; 41.30-53.03 with moderate, mild, and severe anemia accounting for 45.11, 42.86, and 12.03%, respectively. The majority of the anemia (94%) was of the normocytic normochromic type. It was associated with postpartum hemorrhage (AOR = 2.23; 95% CI: 1.24-4.01), cesarean section (AOR = 4.10; 95% CI: 2.11-7.78), lack of iron and folate supplementation during pregnancy (AOR = 2.12; 95% CI: 1.17-4.02), and low diet diversity level (AOR = 1.83; 95% CI: 1.05-3.18). The prevalence of anemia was found to be a major public health concern. Iron and folate supplementation during pregnancy, improved management of PPH, an effective cesarean section with post-operative care, and taking a diversified diet will reduce the burden. Therefore, identified factors should be considered to prevent and control postpartum anemia.

Platelet Satellitism

Platelet satellitism is a rare in vitro phenomenon presenting with platelets rosetting around neutrophils observed in peripheral blood smears prepared from blood samples anticoagulated with ethylenediaminetetraacetic acid (EDTA) but not with heparin sample. The subject was a 65-year-old woman presenting with pain abdomen at our hospital. As a routine investigation, complete blood count (CBC) was advised, and collection was done in EDTA vial. The sample was run in Sysmex Xs800i five-part hematology cell counter. The results of CBC were as follows: total leucocyte count: 15.59 × 103/uL; red blood cell count: 4.61 × 106/uL; hemoglobin: 12.8 g/dL; packed cell volume: 39.5%; mean corpuscular volume: 85.7fl; mean corpuscular hemoglobin (MCH): 27.8 pg; MCH concentration: 32.4 g/dL; and platelet: 26 × 103/uL. On preparation of blood smear and after staining with Leishman stain, the smear showed the formation of platelet rosette around the neutrophils. Platelet satellitism is, hence, defined as the formation of platelet rosetting or clumping around the neutrophils in sample collected in EDTA. Microscopic examination of samples collected in EDTA vial is important before commenting on platelet count.

Hematologi dan Histopatologi Insang Ikan Lele Hasil Budidaya Pembudidaya Lokal di Noekele, Kabupaten Kupang Timur

The aim of this study was to determine the haematological and histopathological of the Catfish gills cultured in Noekele, East Kupang, East Nusa Tenggara. The fish samples used were catfish taken randomly as many as 80 individuals from the rearing pond owned by local farmers in Noekele, then taken to the laboratory for measurement of length and weight, morphological abnormalities observation, preparation of fish blood smears and collection of gills for histological preparation. The observation of morphological abnormalities showed body lesion (90%), one ventral fins (31.3%), one pectoral fins (28.8%), dull and pale body colors (100%). Erythrocyte cell damage on haematological observation on blood smear were tear drop shaped, fusion, lacerated membrane, nuclear extrusion, blebbed nuclei, sperocytes (deformed cells), lysis and shrinking erythrocytes. Based on histopathological analysis, gill’s damage were secondary lamella edema, primary lamella vacuoles, secondary lamella necrosis, epithelium lifting, secondary lamella fusion. Loss of lamella structure, hyperplasia and presence of parasites.

Bovine cerebral theileriosis: first molecular report in cross bred cattle calf in India.

Bovine tropical theileriosis caused by Theileria annulata, is a serious constraint to Indian dairy industry with more fatal infections in exotic cattle and substantial losses to cross-bred and indigenous zebu cattle. The present communication is to place on record the first report of molecular based confirmed case of cerebral theileriosis caused by T. annulata coupled with its morphological detection, clinical manifestations, haematological alterations and therapeutic management in a cross bred cattle calf from India. After preparation of peripheral thin blood smear from cross bred cattle calf at the site of collection and fixation with methanol, blood sample brought to Department of Veterinary Parasitology, College of Veterinary Science and A.H, Jabalpur and stained by standard protocol for Giemsa staining. Genomic DNA was isolated from the collected blood sample using QIAamp® DNA blood mini kit following the manufacturer's recommendations and PCR was performed. The cross bred cow calf revealed high rise in temperature (105.5°F), increased heart rate, labored breathing with seromucous nasal discharge, enlargement of prescapular lymph node and animal exhibited tonic clonic convulsions in response to any sudden noise. Giemsa stained thin blood smear revealed intraerythrocytic piroplasm and Koch'sblue bodies of T. annulata within the cytoplasm of lymphocytes. The species of Theileria was confirmed by molecular amplification of genomic DNA as T. annulata.

FRUIT PEEL EXTRACT OF Sygyzium cumini AS AN ALTERNATIVE STAINING FOR PERIPHERAL BLOOD SMEAR PREPARATIONS

Peripheral Blood Smear is preparations of human blood cells made by smear method and through a staining process in hematological examination to see the elements of peripheral blood cells, namely erythrocytes, leukocytes and platelets. The result of the Peripheral Blood Smear examination is the quality of the staining. This study was conducted with the aim of observing blood cells in blood smear preparations using aqueous extract from the skin of the jamblang fruit (Sygyzium cumini) as an alternative dye to replace Giemsa dye. This research was conducted by comparing the results of erythrocytes, leukocytes and platelets with Giemsa staining and using an alternative staining of fruit peel extract. The sample in this study used blood, and then it was made as a thin blood smear. The results of this study showed that only erythrocytes had a slightly good staining quality while platelets and leukocytes were not good because the acid content in Sygyzium cumini extract could not bind the cell nucleus in platelets and leukocytes. The morphology of normal erythrocyte cells obtained was round and oval disc-like (biconcave) with a purple center and pale edges.

Pengaruh Variasi Waktu Fiksasi Sediaan Apus Darah Tepi (SADT) pada Pengecatan Giemsa terhadap Morfologi Sel Darah Merah

The Blood Smear is a slide whose one of the surfaces is coated using a thin layer of blood and stained with painting of giemsa or wright. Before painting, the preparation is fixated with methanol (methyl alcohol). Prolonged fixation should be avoided because methanol affects subsequent painting and causes some inhibition of intracellular enzymes. Methanol that is left too long in the open air will evaporate (decrease in concentration) and contain water so that it will affect erythrocyte morphology. This study aims to find out the effect of variations in sadt fixation time on giemsa painting on the morphology of red blood cells. This type of research is an experiment. The sample numbered 30 in the form of peripheral blood smear preparations divided by 3 based on variations in fixation time of 3 minutes, 5 minutes and 10 minutes. The results of the Chi-Square statistical test obtained a signification value of 0.00 &lt; 0.05 means that there is a significant difference in time variation in the morphological quality of cells. And the fixation time is best with a fixation time of 3 minutes, at a fixation of 5 minutes still found some crenation cells (cell grinding) with moderate criteria (&lt;3) while fixation with a time of 10 minutes is found a lot of crenation cells (&gt;3).

Serologic evidence of selected vector-borne pathogens in non-owned dogs in the southeast US.

Vector-borne pathogens (VBP) associated with ectoparasitism are of concern for animal health, and there are many gaps in surveillance and reporting data. The purpose of this study was to test for four VBPs in a subset of non-owned dogs from county humane societies in Alabama and Georgia that were admitted to the Auburn University College of Veterinary Medicine Hoerlein Spay/Neuter Program for health exams and routine procedures, including bloodwork and testing with the 4Dx® SNAP® Plus (IDEXX Laboratories, Inc., Westbrook, Maine). Visualized ectoparasites were noted and preserved for identification and analysis. From May-October 2019, residual blood (n=114) was used for preparing blood smears and DNA extraction and PCR. Out of 114 samples, 35.1% (40/114) were seropositive for one or more VBP: Dirofilaria immitis antigen (20.2%; 23/114) and Ehrlichia spp. antibodies (20.2%; 23/114); six VBD-positive dogs (15%) tested positive for both. No dogs had detectable antibodies to Borrelia burgdorferi or Anaplasma spp. (0%; 0/114). Microfilariae of D. immitis were present in 7 blood smears, all from dogs that were D. immitis antigen positive. Morulae or DNA of Ehrlichia or Anaplasma spp. were not identified in any sample. Fleas were documented in 20.4% (23/113) of dogs, 9.7% (11/113) were infested with ticks, predominantly Amblyomma americanum, and co-infestations were noted in 2.7% (3/113). Our data indicate that there is substantial VBP risk in Alabama and Georgia, and that the reservoir potential of domestic animals, especially non-owned animals, along with potential wildlife reservoirs warrants further investigation.

Autohaem: 3D printed devices for automated preparation of blood smears.

The process of making blood smears is common in both research and clinical settings for investigating the health of blood cells and the presence of blood-borne parasites. It is very often carried out manually. We focus here on smears for malaria diagnosis and research, which are frequently analyzed by optical microscopy and require a high quality. Automating the smear preparation promises to increase throughput and to improve the quality and consistency of the smears. We present here two devices (manual and motorized) designed to aid in the making of blood smears. These are fully documented, open-source hardware, and an important principle was to make them easily fabricated locally anywhere. Designs and assembly instructions are freely available under an open license. We also describe an image analysis pipeline for characterizing the quality of smears and use it to optimize the settings and tunable parameters in the two devices. The devices perform as well as expert human operators while not requiring a trained operator and offering potential advantages in reproducibility and standardization across facilities.