Smear Examination Research Articles

Epidemiological situation of bovine tropical theileriosis in an arid region in central Tunisia with a phylogenetic analysis of Theileria annulata.

Tropical theileriosis, Theileria annulata infection, is the most prevalent summer disease in Tunisia. It is transmitted by Hyalomma scupense, a two-host tick known to be endophilic. The present study aimed to estimate the infection prevalence of cattle by T. annulata in two districts from central Tunisia. Blood samples collected from 270 Holstein cattle from the Sidi Bouzid (140 samples) and Kasserine districts (130 samples) were analysed by Giemsa staining and T. annulata-specific PCR. In both regions, PCR revealed a prevalence of 32.6%. This was significantly higher than the 6.3% prevalence obtained by Giemsa staining blood smears (p < 0.001). Giemsa staining also revealed a low parasitaemia of 0.05%. The PCR-based prevalence was not statistically different between the two districts (31.4 ± 0.04 and 33.8 ± 0.04% in Sidi Bouzid and Kasserine districts, respectively, p = 0.6). On the contrary, the results of blood smear examination (2.85 and 10% in Sidi Bouzid and Kasserine, respectively) differed significantly between the two sampling sites (p = 0.01). There was no evidence of a statistically significant difference between the overall molecular infection prevalence when the samples were segregated based on animals' age or gender (p = 0.1 and 0.2, respectively) and a similar trend was observed for Giemsa staining. Ten PCR amplicons of the Tams1 gene (721bp) were subsequently sequenced from the two regions. The phylogenetic analyses showed 100% similarity between all sequences. The unique conserved Tams1 sequence was deposited in GenBank under the accession number OP428816 and used to infer its phylogenetic relationships with those available in the GenBank repository. This is the first report of the presence of T. annulata in this region of central Tunisia which has no history of tropical theileriosis. Priority areas for future studies include understanding the origin of these T. annulata-positive animals in a region where the presence of a known natural vector tick, H. scupense, has not been reported. Given that the disease severely constrains cattle productivity, it would also be worthwhile to investigate if other potential vectors for T. annulata, such as Hyalomma dromedarii, are present in the arid regions.

Variations in complete blood counts and cellular morphological alterations in blood smear of COVID-19 patients from a tertiary care center

Corona viruses are a large family of viruses of which Corona virus disease 19 (COVID 19) infection evokes a hyperinflammatory response and severe acute respiratory distress syndrome in human beings. Complete blood count evaluation and peripheral smear examination is an essential tool for early diagnosis and management of COVID-19 positive patients. This study was aimed to evaluate the variations in complete blood counts (CBC) and changes in the cellular morphology in the blood smear in patients affected with COVID-19. This was an observational study, undertaken in a tertiary care center. The study included baseline blood samples from 96 proven COVID-19 positive cases. The samples were subjected for automated complete blood count analysis and peripheral smear examination.Thequantitative data were expressed by mean and Standard deviation. Qualitative categorical variables were expressed as numbers and percentages. Out of the total 96 cases, the most common quantitative finding was neutrophilic leukocytosis with relative lymphopenia. Five cases of absolute lymphopenia was observed. Majority of the cases had normal hemoglobin values and normal platelet counts. The characteristic morphological changes were seen in neutrophils and lymphocytes. The neutrophils showed presence of toxic granules, cytoplasmic vacuolations and hypolobation.The morphological alteration in lymphocytes included reactive lymphocytes with plasmacytoid and monocytoid morphology, large granular lymphocytes and Turk cells. Infrequent finding of leukoerythroblastic reaction was also observed in our study.To summarize, hematological investigation of COVID 19 cases may show several abnormal findings. Quantitative findings like Neutrophilia and lymphopenia were characteristic, whereas, the most common morphological alteration observed was atypical lymphocytes. Quantitative and qualitative alterations of hematological parameters are one of the investigations which would aid timely diagnosis of the disease. Toxic granules and cytoplasmic vacuolations in neutrophils may indicate ongoing sepsis in COVID 19 patients which in turn aids in efficient management of the patients to reduce the mortality rate.

Iron Deficiency Anemia Presenting With Pancytopenia: A Study From India.

Iron deficiency anemia (IDA) is the most common cause of anemia worldwide. IDA is commonly associated with thrombocytosis and normal or slightly decreased leukocyte count. Sometimes it can present with thrombocytopenia, but rarely present with pancytopenia. Here we are presenting six cases of severe iron deficiency presenting with pancytopenia, which responded to iron replenishment. This 12-month observational study was conducted in the Department of General Medicine at a tertiary care Centre in India. All cases of pancytopenia (after exclusion of other causes) with IDA were included. IDA was established with the help of a complete blood count (CBC), peripheral smear examination, serum iron studies, and serum ferritin. Results: In our study, CBC at four weeks later of iron transfusion without other supplementation showed significant improvement in hematological parameters. Severeiron deficiency is a reversible etiology of pancytopenia. It should be kept as a differential diagnosis of pancytopeniaif common causes of pancytopenia are ruledout.

Clinical, Haemato-biochemical Evaluation and Comparison of Polymerase chain reaction with Conventional Blood Smear Examination for the Detection of Babesia gibsoni Infection in Dogs

The objective of the present study was to detect Babesia gibsoni infection in dogs which are presented to Veterinary Clinical Complex, Rajiv Gandhi Institute of Veterinary Education and Research, Puducherry. A total of 85 dogs with clinical signs such as inappetence, fever, enlarged lymph nodes and tick infestation were screened. The diagnostic efficacy of polymerase chain reaction (PCR) technique has been compared with blood smear examination to detect B. gibsoni infection in dogs. By blood smear and PCR examination, six (7.1%) dogs and 28 (42.4%) dogs were positive for B. gibsoni infection respectively. Clinical examination of dogs with B. gibsoni showed tick infestation, anorexia, dullness, pyrexia, pale/congested conjunctival mucus membrane, tachypnoea and swollen lymph nodes. In addition, few dogs had icterus and haemoglobinuria. Laboratory examination of blood samples of B. gibsoni dogs revealed significant reduction in haemoglobin, PCV, erythrocyte count, platelet count and serum albumin levels. Significant increase in levels of ALP, globulin, bilirubin, BUN and creatinine were recorded.

Systemic manifestation of necrotic erythema nodosum leprosum: A case report of a fatal leprosy.

Necrotic erythema nodosum leprosum (ENL) is an uncommon manifestation of type 2 lepra reaction, encountered in lepromatous and borderline lepromatous cases of leprosy. Necrotic ENL is associated with the involvement of multiple organs, therefore delayed diagnosis and treatment will lead to complications and poor prognosis. The aim of this case report was to report a challenging case of necrotic ENL misdiagnosed with multiple cellulitis since there were no signs of prior leprosy nor had any antimycobacterial treatment. A 45-year-old man was presented to the surgery department of Dr. Zainoel Abidin Hospital, Banda Aceh, Indonesia, with complaints of fever, joint pain, and painful tender skin lesions with ulceration over the trunk, extremities, and ears for one month. The patient was diagnosed clinically with multiple cellulitis and underwent a debridement procedure. Clinical improvement was absent, the patient was then consulted to the dermatology department. Physical examination showed normal vital signs, madarosis, inguinal lymphadenopathy, thickening of nerves, and sensation of numbness in both hands and feet. Laboratory examinations on admission showed leucocytosis, anemia, thrombocytopenia, hypoalbuminemia, hypocalcemia, and elevated creatinine and ureum level. A slit skin smears examination yielded positive acid-fast bacilli (AFB) with a bacteriological index (BI) value of 3+ and morphological index (MI) of 72%. The patient was diagnosed with lepromatous leprosy with necrotic ENL reaction. Intravenous methylprednisolone and cefoperazone-sulbactam were given. Multidrug therapy mulitbacillary (MDT-MB) without dapsone, and ofloxacin 400 mg was initiated. On day 17, the patient had septic shock. The patient became unconscious and experienced death. This case highlights that medical professionals should be aware of the various manifestations of necrotic ENL to correctly diagnose and provide treatment as soon as possible to prevent mortality, especially in leprosy-endemic country, Indonesia.

Utility of VCS Parameters as a Cost-effective and Early Marker of Sepsis: A Hospital-based Study.

Sepsis is a life-threatening condition. Nowadays, hospitals rely on laboratory parameters like CRP and procalcitonin to detect sepsis. There is a need to evaluate and validate more accurate and early predictors of sepsis in critically ill patients. We analyzed volume, conductivity, and scatter (VCS) parameters of leukocytes in sepsis patients and compared them with the control group with no illness. It was a prospective case-control study. A total of 80 patients were studied with 40 sepsis cases and 40 controls. Peripheral smear examination was done in all the cases. Other parameters, such as WBC count, neutrophil%, absolute neutrophil count (ANC), High-sensitive C-reactive protein (hs-CRP), procalcitonin, and blood cultures were analyzed. We took the data of the patients from medical records and correlated it with other tests. Complete blood picture reports were generated by the Beckman Coulter LH series (LH 750 and 780). VCS parameters for neutrophils, lymphocytes and monocytes were compared between both groups. The results were analyzed using SPSS software (16.0 version). The age group was 20-85 years with male predominance. The mean neutrophil volume (MNV) and mean monocyte volume (MMV) were higher in the sepsis group when compared with the control group. The mean neutrophil conductivity and scatter was lower in the sepsis group and comparatively higher in the control group, but it was insignificant. The mean neutrophil volume values were higher in the sepsis group even with low total leukocyte count (TLC) when the patient had sepsis. There is no change in the mean lymphocyte VCS parameter and mean monocyte conductivity, scatter in both the groups. The mean neutrophil volume and MMV are found to be good sensitive markers in the prediction of sepsis. Even when TLC is on the lower side in patients with sepsis, these parameters predict sepsis accurately. This helps clinicians to assess sepsis in patients at an early stage and has an important practical implication. Goyal H, Singhal A, Joseph M. Utility of VCS Parameters as a Cost-effective and Early Marker of Sepsis: A Hospital-based Study. Indian J Crit Care Med 2023;27(9):647-650.

FUNGAL DERMATITIS IN LOCAL CAT

Dermatitis is a skin disorder caused by various agents such as fungi, ectoparasites, bacteria and metabolic diseases. The purpose of this research is to find out the causative agent of the infection and how the treatment progresses. The case animal is a male local cat, one and a half months old and 1 kg body weight. came to the Teaching Animal Hospital, Udayana University with weakness and experienced itching that had lasted for 5 weeks. On clinical examination, the cat experienced alopecia and erythema on the ears, feet and stomach. Tape smear examination found spores, Fungal culture found Penicillium Sp., Aspergillus Sp. Staphylococcus sp was found in bacterial culture. Routine blood tests showed that the case cat had normochromic microcytic anemia. Based on the anamnesis, clinical examination, and supporting examinations, it can be concluded that the case cat was diagnosed with dermatitis due to a fungal infection. The prognosis for case cats is fausta. The therapy given was grisofulvin, chlorphenamine maleate, sebasol fish oil shampoo. The results of therapy for two weeks showed that the case cat's condition was getting better with hair growth and reduced frequency of scratching and redness of the skin. Pay attention to the cleanliness of the animal's body and the place of maintenance. Animals infected with the fungus are placed in separate cages during treatment to prevent re-infection.

A rare case of Elizabethkingia meningoseptica infection in a neonate

Background: Elizabethkingia infections were reportedly rare, but if it was found, it had been known to cause neonatal meningitis, bloodstream infections and respiratory infections. Elizabethkingia meningoseptica had a unique antibiotic susceptibility pattern, usually resistant to most antibiotics. Elizabethkingia infections were associated with a high mortality rate because of the lack of effective therapeutic regimens, antibiotic resistance and virulence. Case Presentation: Fourteen days old boy patient came with the chief complaint of seizure, which occurred twice. Before the seizure, the patient had a fever with the highest temperature was 39°C. The patient looked lethargic, tended to sleep more often, cried occasionally and was not as active as previously. Septic marker revealed an Immature to Total neutrophil (IT) ratio of 0.2 and C-Reactive Protein (CRP) 49.30 mg/dL. A blood smear examination showed toxic granules, vacuolization of the leucocyte and reactive thrombocytosis. Cerebrospinal fluid analysis revealed a cell number of 2520 cell/uL, polymorphonuclear (PMN) cell 80%, mononuclear (MN) cell 20%, Nonne and Pandy was positive, protein level at 300 mg/dL and cerebrospinal fluid glucose level below 20 mg/dL. The patient was initially diagnosed with sepsis and meningitis and was given ampicillin and gentamicin. Blood and cerebrospinal fluid culture isolated Elizabethkingia meningoseptica and significantly as the infection-causing agent. The antibiotic sensitivity of the blood culture was ciprofloxacin and levofloxacin. The antibiotic sensitivity of the cerebrospinal fluid culture was levofloxacin. During the treatment, there were several inappropriate results between clinical manifestation and laboratory results; the antibiotic was changed to levofloxacin according to the sensitivity of the blood and cerebrospinal fluid culture. Conclusion: Elizabethkingia meningoseptica was a rare case, and the nature of this bacteria was resistant to multiple antibiotics. Treatment should be considered carefully.

Another Chanarin-Dorfman Syndrome (CDS) in an Egyptian Patient

Abstract Introduction Chanarin–Dorfman syndrome is a very rare autosomal recessive syndrome caused by mutation in ABHD5 a cofactor for adipose triglyceride lipase resulting in intracellular accumulation of triacylglycerol in various body tissues. Several patients were previously reported from Egypt and middle East but still the diagnosis is missed for several years. Here we report another case with this syndrome and describe his journey to diagnosis. Case report A 4.4 year old boy, the fifth in order of birth of first cousin Egyptian parents presented to the Genetics Clinic because of abnormal scaling all over the body since birth. On examination, he had short stature, hepatomegaly, and generalized ichthyosis. His chest, heart and neurological examination showed no abnormalities. Ocular examination revealed refractive errors. Audiometry revealed hearing loss. Peripheral blood smear examination demonstrated cytoplasmic vacuoles within the neutrophils (Jordans bodies ) confirming the diagnosis of CDS. He had family history of sibling death at age of 2 day with extensive scaling overall the body. Conclusion CDS should be suspected in patients with congenital ichthyosis and hepatomegaly. Diagnosis is simple using blood smear to find Jordans bodies.

The potential for zoonotic malaria transmission in five areas of Indonesia inhabited by non-human primates

BackgroundIndonesia is home to many species of non-human primates (NHPs). Deforestation, which is still ongoing in Indonesia, has substantially reduced the habitat of NHPs in the republic. This has led to an intensification of interactions between NHPs and humans, which opens up the possibility of pathogen spillover. The aim of the present study was to determine the prevalence of malarial parasite infections in NHPs in five provinces of Indonesia in 2022. Species of the genus Anopheles that can potentially transmit malarial pathogens to humans were also investigated.MethodsAn epidemiological survey was conducted by capturing NHPs in traps installed in several localities in the five provinces, including in the surroundings of a wildlife sanctuary. Blood samples were drawn aseptically after the NHPs had been anesthetized; the animals were released after examination. Blood smears were prepared on glass slides, and dried blood spot tests on filter paper. Infections with Plasmodium spp. were determined morphologically from the blood smears, which were stained with Giemsa solution, and molecularly through polymerase chain reaction and DNA sequencing using rplU oligonucleotides. The NHPs were identified to species level by using the mitochondrial cytochrome c oxidase subunit I gene and the internal transcribed spacer 2 gene as barcoding DNA markers. Mosquito surveillance included the collection of larvae from breeding sites and that of adults through the human landing catch (HLC) method together with light traps.ResultsAnalysis of the DNA extracted from the dried blood spot tests of the 110 captured NHPs revealed that 50% were positive for Plasmodium, namely Plasmodium cynomolgi, Plasmodium coatneyi, Plasmodium inui, Plasmodium knowlesi and Plasmodium sp. Prevalence determined by microscopic examination of the blood smears was 42%. Species of the primate genus Macaca and family Hylobatidae were identified by molecular analysis. The most common mosquito breeding sites were ditches, puddles and natural ponds. Some of the Anopheles letifer captured through HLC carried sporozoites of malaria parasites that can cause the disease in primates.ConclusionsThe prevalence of malaria in the NHPs was high. Anopheles letifer, a potential vector of zoonotic malaria, was identified following its collection in Central Kalimantan by the HLC method. In sum, the potential for the transmission of zoonotic malaria in several regions of Indonesia is immense.Graphical

Effect of antibiotic therapy of genital mycoplasmosis in pregnant cows on cytological and immunological characteristics of vaginal secretions

Purpose: the aim is to study the therapeutic efficacy of the macrolide group antibiotic tulatromycin and its effect on the immunological and cytological characteristics of vaginal secretions.Materials and methods. For the experiment, 2 groups of 8 animals each were formed. The first group - cows without clinical signs of vaginitis with a positive PCR test for Mycoplasma spp.; the second group - clinically healthy cows with a negative PCR test for Mycoplasma spp. The first group of cows, the group for the treatment of genital mycoplasmosis, used the antibiotic traksovet 100 (tulatromycin) at a dose of 2.5 mg per 1 kg of animal body weight, subcutaneously, once 40 days before the expected delivery. Vaginal secretions were collected from both groups of cows, in which the content of Ig G, Ig M, Ig A, sIg A, lysozyme activity and cytological examination of the vaginal smear were determined.Results. The use of tulatromycin in cows with genital mycoplasmosis leads to a decrease in the content of Ig M and sIg A in vaginal secretions, which reach the values of these indicators in healthy animals. A less pronounced positive effect is observed with respect to the activity of the vaginal secretion lysozyme, which, although significantly increased, remains lower than in healthy cows. Cytological examination of vaginal smears against the background of antibiotic therapy showed a significant decrease in the number of epithelium and leukocytes.Conclusion. The use of traksovet 100 antibiotic as etiotropic therapy for cows with genital mycoplasmosis leads to the elimination of the pathogen from the vaginal mucosa in 75 % of sick animals and provides partial restoration of cytological and immunological characteristics of vaginal secretions.

Comparing MitoQ10 and heat therapy: Evaluating mechanisms and therapeutic potential for polycystic ovary syndrome induced by circadian rhythm disruption

ABSTRACT Environmental factors, such as sleep restriction, contribute to polycystic ovary syndrome (PCOS) by causing hyperinsulinemia, hyperandrogenism, insulin resistance, and oligo- or anovulation. This study aimed to evaluate the effects of circadian rhythm disruption on reproductive and metabolic functions and investigate the potential therapeutic benefits of MitoQ10 and hot tub therapy (HTT). Sixty female rats were divided into six groups: control, MitoQ10, HTT, and three groups with PCOS induced by continuous light exposure(L/L). The reproductive, endocrine, and structural manifestations ofL/L-induced PCOS were confirmed by serum biochemical measurements, ultrasound evaluation of ovarian size, and vaginal smear examination at week 14. Subsequently, the rats were divided into the L/L (untreated), L/L+MitoQ10-treated, andL/L+HTT-treated groups. At the end of week 22, all rats were sacrificed. Treatmentwith MitoQ10 or HTT partially reversed the reproductive, endocrine, and structural features of PCOS, leading to a decreased amplitude of isolated uterine contractions, ovarian cystic changes and size, and endometrial thickness. Furthermore, both interventions improved the elevated serum levels of anti-Mullerian hormone (AMH), kisspeptin, Fibulin-1, A disintegrin and metalloproteinase with thrombospondin motifs 19 (ADAMTS-19), lipid profile, homeostatic model assessment for insulin resistance (HOMA-IR), oxidative stress markers, androgen receptors (AR) and their transcription target genes, FKBP52 immunostaining in ovarian tissues, and uterine estrogen receptor alpha (ER-α) and PRimmunostaining. In conclusion, MitoQ10 supplementation and HTT demonstrated the potential for ameliorating metabolic, reproductive, and structural perturbations associated with PCOS induced by circadian rhythm disruption. These findings suggest a potential therapeutic role for these interventions in managing PCOS in women.

Development of a vertical bubble acoustic microfluidic sputum liquefier for improved cancer cell detection

The detection of cancer in the lower respiratory tract is often performed using sputum smears, which is a convenient method. However, due to its high viscosity, it can be difficult to concentrate target cancer cells from sputum. To improve sputum smear and cancer cell detection, it is highly desirable to have efficient sputum liquefaction approaches. Our study focused on the development of a vertical bubble acoustic microfluidic sputum liquefier that can mix sputum and lysis buffer in a harmless and efficient manner. The system can automatically process sputum samples with a wide range of viscosities without vortexing. Moreover, well-liquefied sputum samples can easily concentrate cancer cells, leading to improved efficiency and accuracy of diagnosis compared to the direct smear method. We expect that our device will increase the positive rate of pathological examination of sputum smears in patients with lung cancer in the future.

A single Haemoproteus plataleae haplotype is widespread in white ibis (Eudocimus albus) from urban and rural sites in southern Florida.

The American white ibis (Eudocimus albus), a common bird species in Florida, has become increasingly urban, with many populations relying heavily on urban and suburban habitats, which may alter parasite transmission. Parasites of ibis, especially haemosporidians, are understudied. Avian haemosporidia can have a wide range of impacts on birds, including decreased reproductive success or increased mortality. Because southern Florida is subtropical and has a high diversity of potential vectors for haemosporidia, we hypothesized that there will be a high prevalence and genetic diversity of haemosporidia in white ibis. A total of 636 ibis from South Florida were sampled from 2010 to 2022, and blood samples were tested for haemosporidia by examination of Giemsa-stained thin blood smears and/or nested PCRs targeting the cytochrome b gene. A total of 400 (62.9%, 95% CI 59-66.7%) ibis were positive for parasites that were morphologically identified as Haemoproteus plataleae. Sequences of 302 positives revealed a single haplotype of Haemoproteus (EUDRUB01), which was previously reported from white ibis in South Florida and captive scarlet ibis (E. ruber) in Brazil. No Plasmodium or Leucocytozoon infections were detected. Parasitemias of the 400 positive birds were very low (average 0.084%, range 0.001%-2.16% [although only 2 birds had parasitemias >1%]). Prevalence and parasitemias were similar for males and females (68% vs. 61.6% and 0.081% vs. 0.071%, respectively). Prevalence in juveniles was lower compared with adults (52% vs. 67.4%) but parasitemias were higher in juveniles (0.117% vs. 0.065%). This data shows that H. plataleae is common in ibis in South Florida. Although parasitemias were generally low, additional research is needed to determine if this parasite has subclinical effects on ibis, if additional haplotypes or parasite species infect ibis in other regions of their range, or if H. plataleae is pathogenic for other sympatric avian species.

Rapid and Ultrasensitive Detection of Plasmodium spp. Parasites via the RPA-CRISPR/Cas12a Platform.

Microscopic examination of thick and thin blood smears stained with Giemsa dye is considered the primary diagnostic tool for the confirmation and management of suspected clinical malaria. However, detecting gametocytes is relatively insensitive, particularly in asymptomatic individuals with low-density Plasmodium infections. To complement existing diagnostic methods, a rapid and ultrasensitive point-of-care testing (POCT) platform for malaria detection is urgently needed and necessary. A platform based on recombinase polymerase amplification (RPA) followed by CRISPR/Cas12a (referred to as RPA-CRISPR/Cas12a) was developed and optimized for the determination of Plasmodium spp. parasites, particularly Plasmodium falciparum, using a fluorescence-based assay (FBDA), lateral flow test strips (LFTS), or naked eye observation (NEO). Then, the established platform was assessed with clinical malaria isolates. Under optimal conditions, the detection threshold was 1 copy/μL for the plasmid, and the limit of detection was 3.11-7.27 parasites/μL for dried blood spots. There was no cross-reactivity against blood-borne pathogens. For the accuracies of RPA-CRISPR/Cas12a, Plasmodium spp. and P. falciparum testing were 98.68 and 94.74%, respectively. The method was consistent with nested PCR results and superior to the qPCR results. RPA-CRISPR/Cas12a is a rapid, ultrasensitive, and reliable platform for malaria diagnosis. The platform requires no or minimal instrumentation for nucleic acid amplification reactions and can be read with the naked eye. Compared with similar diagnostic methods, this platform improves the reaction speed while reducing detection requirements. Therefore, this platform has the potential to become a true POCT for malaria parasites.

Cattle co-infection patterns by hemopathogens and their phylogenetic analysis during the tick season in Constantine and Mila, Northeast Algeria.

Tropical theileriosis, babesiosis, and anaplasmosis are the most dominant tick-borne infections in North Africa where they cause significant economic losses in ruminants' industry. The aim of the present work was to study infections and co-infection patterns in 66 cattle with clinical signs of piroplasmosis and/or anaplasmosis in two localities, Beni Hamidene and Grarem Gouga, districts of Constantine and Mila (Northeast of Algeria), respectively. This study was conducted between early May and late September during four years 2017, 2018, 2020, and 2021. PCR showed that the most frequent pathogen in cattle with clinical signs of piroplasmosis and/or anaplasmosis was Theileria annulata (66/66; 100%) followed by Babesia bovis (21/66; 31.8%), Anaplasma marginale (15/66; 22.7%), and Babesia bigemina (3/66; 4.5%) (p < 0.001). Giemsa-stained blood smears examinations revealed that 66.7% (44/66); 10.6% (7/66); and 9.1% (6/66) of cattle were infected by T. annulata, Babesia spp., and A. marginale, respectively (p < 0.001). PCR revealed seven co-infection patterns: T. annulata/A. marginale (15/66; 22.7%), T. annulata/B. bovis (21/66; 31.8%), T. annulata/B. bigemina (3/66; 4.5%), T. annulata/A. marginale/B. bovis (7/66; 10.6%), T. annulata/B. bovis/B. bigemina (2/66; 3%), T. annulata/A. marginale/B. bigemina (1/66; 1.5%), and T. annulata/A. marginale/B. bigemina/B. bovis (1/66; 1.5%). Phylogenetic analyses showed that T. annulata Tams1 and B. bigemina gp45 sequences were identical to isolates from Mauritania and South Africa, respectively. The three A. marginale amplicons obtained herein had 99.63 to 99.88% similarity between them. This study provides data that can be used to improve control programs targeting these cattle hemopathogens.

A high-resolution large-scale dataset of pathological and normal white blood cells

Microscopic examination plays a significant role in the initial screening for a variety of hematological, as well as non-hematological, diagnoses. Microscopic blood smear examination that is considered a key diagnostic technique, is in recent clinical practice still performed manually, which is not only time consuming, but can lead to human errors. Although automated and semi-automated systems have been developed in recent years, their high purchasing and maintenance costs make them unaffordable for many medical institutions. Even though much research has been conducted lately to explore more accurate and feasible solutions, most researchers had to deal with a lack of medical data. To address the lack of large-scale databases in this field, we created a high-resolution dataset containing a total of 16027 annotated white blood cells. Moreover, the dataset covers overall 9 types of white blood cells, including clinically significant pathological findings. Since we used high-quality acquisition equipment, the dataset provides one of the highest quality images of blood cells, achieving an approximate resolution of 42 pixels per 1 μm.

Effect of pathological high shear exposure time on platelet activation and aggregation.

Circulating platelets are sometimes exposed to high shear rate environments due to vascular stenosis, and the effect of transiently elevated pathological high shear rates on platelet activation and aggregation function has not been clarified. The aim of this study was to investigate the effect of pathological high shear rate (8302s-1) exposure time (3.16-25.3 ms) on platelet activation and aggregation function. In addition, by adding active ingredients of antiplatelet drugs such as ASA (an active ingredient of aspirin), Ticagrelor, Tirofiban and GP1BA (platelet membrane protein GPIb inhibitor) in vitro, we studied TXA2, P2Y12-ADP, GPIIb/IIIa-fibrinogen and GPIb /IX/V-vWF receptor pathways to determine platelet activation function mediated by pathological high shear rate. In this study, we designed a set of microfluidic chips with stenosis lengths of 0.5 mm, 1 mm, 2 mm, 3 mm, and 4 mm, all with 80% stenosis, to generate pathological high shear forces that can act at different times. The whole blood flowing through the microchannels was collected by perfusion of sodium citrate anticoagulated whole blood at a physiological arterial shear rate (1500 s-1), and the expression levels of platelet surface activation markers (P-selectin and GP IIb/IIIa) and the degree of platelet aggregation were analyzed by flow cytometry; platelet aggregation patterns were observed by microscopic examination of blood smears. The results showed that shearing significantly increased platelet activation and aggregation levels compared to un-sheared whole blood, and the activation and aggregation levels increased with increasing duration of pathological high shear rate. In vitro inhibition studies showed that ASA barely inhibited the expression of P-selectin and PAC-1 on the platelet surface; Ticagrelor effectively inhibited the expression of both P-selectin and PAC-1; Tirofiban significantly inhibited the expression of PAC-1 on the platelet surface and slightly inhibited the expression of P-selectin; GP1BA significantly inhibited the expression of both. Our results suggest that transient pathological high shear rate (8302s-1) exposure can induce platelet activation in a time-dependent manner; however, the mechanism is more complex and may be due to the following reasons: transient elevated pathological high shear rate activates platelets through the GPIb/IX/V-vWF receptor pathway, and after platelet activation, its surface membrane protein GPIIb/IIIa receptors activate platelets through fibrinogen to form platelet-platelet aggregates, and further activation of active substances such as ADP and TXA2 released by platelet alpha particles, which contribute to the formation of irreversible platelet aggregation.

Cryptosporidiosis in HIV-seronegative patients: A case series

Enteric infections by are a frequent cause of diarrhea among immunocompromised and HIV infected patients but there are limited reports of their occurrence in immunocompetent patients. Cryptosporidiosis is a faeco-orally transmitted protozoan infection and is one of the leading causes of chronic parasitic diarrhea. Here we report a case series of cryptosporidiosis from three different patients where two patients are immunocompetent and one is a post renal transplant case. All the patients came with watery diarrhea for which stool samples were collected and examined by modified Ziehl Neelsen acid fast staining (Kinyoun’s method). On microscopic examination of smear, multiple, pink, round to oval acid fast oocysts of 4-6µm diameter were seen against a blue background morphologically resembling . Early diagnosis and prompt initiation of antiparasitic drugs aids in timely treatment and prevention of disseminated cryptosporidiosis.

Isolation of Malassezia yeasts from dogs with gastrointestinal disease undergoing duodenal endoscopy.

To culture Malassezia and other fungi from the duodenum of dogs with gastrointestinal signs undergoing routine endoscopic examination. Quantitative microbial culture was performed on duodenal juice aspirated from dogs with suspected enteropathy during routine upper gastrointestinal endoscopy; samples were cultured on Sabouraud's dextrose agar (30, 32 and 37°C) and modified Dixon agar (32°C) for 14 days. Isolates were identified phenotypically and by matrix-assisted laser desorption ionisation-time of flight, and internal transcribed spacer sequencing. Yeast presence was also evaluated by cytological and histopathological examination of smears and biopsy specimens. Forty-five dogs were recruited with chronic inflammatory enteropathy (n=38), granulomatous colitis (n=2), gastric adenocarcinoma (n=2), duodenal small cell lymphoma (n=1) and idiopathic severe gastrointestinal haemorrhage (n=2). Fungi were cultured from 14 dogs: Malassezia pachydermatis was isolated from eight [chronic inflammatory enteropathy (n=7) (along with Candida albicans n=1); granulomatous colitis (n=1)] and Malassezia sympodialis from another (gastric adenocarcinoma). Five dogs with chronic inflammatory enteropathy yielded other yeasts (C. albicans, Candida glabrata, Kazachstania slooffiae, Kazachstania telluris, Pichia kudriavzevii [syn. C. krusei]). Yeasts were never observed in histopathological specimens. Fluorescent microscopical examination of cytological specimens showed yeast in only one case, from which K. slooffiae was subsequently isolated. Based on a literature search, this is the first report of isolation of M. pachydermatis, M. sympodialis, K. slooffiae and K. telluris from the canine duodenum. Further studies are needed to determine whether these are resident or transient fungi in the canine duodenum and whether their presence has a pathogenic effect on the host.