Parasitological Tests Research Articles

Gastrointestinal helminth of Nile crocodiles (Crocodylus niloticus), in Arba Minch Crocodile Ranch, Ethiopia

Parasites are among the major organisms that affect the health of crocodiles. A cross-sectional study was conducted from November 2021 to May 2022 aimed to determine the prevalence of gastrointestinal helminths of Nile crocodiles in Arba Minch Crocodile Ranch (AMCR), Ethiopia. A total of 322 fecal samples were collected using a random sampling technique. Standard parasitological tests were used to identify the gastrointestinal helminth parasite. Out of 322 fecal samples examined, 102(31.68%) were found positive for gastrointestinal helminths. Among the endoparasite eggs detected Dujardinascaris (18.01%) species was the most dominant species followed by Trichinella species (5.62%), polydeliphs (3.1%) and Renifers species (0.62%). There was a statistically significant difference(P = 0.0001) in the prevalence of helminth parasites between male (15.97%) and female (44.38%) crocodiles. Likewise, the prevalence of helminths between different age groups was determined with the highest prevalence in crocodiles of less than 8 years old (41.9%) and the lowest prevalence in crocodiles of age greater than 11 years (19.56%). It has been observed that the crocodile in the area of study had mixed infestation with more than one parasite. The study revealed that gastrointestinal parasites were prevalent in the ranch. Therefore, strict prophylaxis treatment and hygiene should be performed in the study area.

The STROGHAT study protocol: An intervention study to evaluate safety, effectiveness and feasibility of treating gambiense HAT seropositive subjects with acoziborole.

Background Coordinated efforts in the control of gambiense human African trypanosomiasis (gHAT) have significantly reduced its endemicity. WHO targets interruption of transmission by 2030. However, challenges remain, including low sensitivity of the current parasitological confirmation tests, leaving a potential human reservoir untreated. Acoziborole, a single-dose oral treatment, effective in both disease stages with a good safety profile, offers the potential of treatment of parasitologically negative gHAT seropositive subjects, which could improve diagnostic sensitivity. The STROGHAT study aims to evaluate whether this approach can lead to elimination of T.b. gambiense from its human reservoir, and to provide further safety data on acoziborole. It also includes a costing analysis and a prospective evaluation of the performance of the screening and diagnostic tests used. Methods STROGHAT is a one-arm epidemiological study, with a nested phase IIIb, one-arm, open label, non-randomized, multicentre clinical trial. It will be implemented over four years in the endemic region of the Equateur North, in the Democratic Republic of Congo. For the first three years, parasitologically negative gHAT seropositive subjects will be treated with acoziborole, while parasitologically confirmed cases will receive standard of care. Individual follow-up needs and accurate prevalence estimate will be based on immunological and molecular tests performed for all gHAT screening test seropositive subjects at a reference laboratory. In the fourth year, standard procedures will resume, and a prevalence survey will assess whether interruption of transmission has been achieved. Discussion The manuscript outlines the study background, objectives and methods while discussing its strengths and challenges. If successful, the STROGHAT study will provide critical evidence on the effectiveness, safety and feasibility of the new strategy, and inform future elimination strategies. Clinical trial registration NCT06356974. Date of registration: April 4, 2024.

Onchocerciasis is not a major cause of blindness in two endemic villages in Sierra Leone

ObjectiveSierra Leone, a country where onchocerciasis is endemic in 14 of the 16 districts, was the focus of our investigation. Despite 17 rounds of annual ivermectin treatment since 2005, a report circulated by a local politician indicated an increase in cases of suspected onchocerciasis-related vision impairment in two villages (Mangobo and Petifu) in Tonkolili district. In response, the National Neglected Tropical Disease Program conducted a comprehensive investigation. Ophthalmological, parasitological, and serological tests were conducted using standard procedures to determine the relationship between self-reported vision loss and onchocerciasis in adults. In addition, serological tests were carried out on children aged 5 to 9 years to assess the recent status of exposure to onchocerciasis in the two villages.ResultsReported vision loss in 37 patients was mainly due to cataracts (35.1%), allergic conjunctivitis (18.9%), refractive error (10.8%), and other conditions not related to onchocerciasis. There were 40.7% of all adults (N = 54) tested and 29.0% of 31 persons with self-reported vision loss who were positive for Ov-16 IgG4 antibodies, suggesting a history of exposure to onchocerciasis. However, otoscopic eye examinations and microscopic skin snip tests were all negative for Onchocerca volvulus microfilariae, indicating no active or low-intensity infection among adults and a low or zero risk of serious ocular morbidity in the two villages. Onchocerciasis may no longer be a major cause of blindness in these two villages. Apparently, 4.6% of 153 children aged 5 to 9 years tested positive for Ov-16 IgG4 antibodies, suggesting that onchocerciasis transmission is likely still ongoing in the two villages. The data presented here suggest that more annual rounds of mass treatment with ivermectin with high coverage are needed to eliminate onchocerciasis transmission in this area.

Congenital Chagas disease: A cohort study to assess molecular diagnostic methods at the Chagas disease national reference center of Argentina.

Trypanosoma cruzi is a protozoan parasite which causes Chagas disease. Mother-to-child transmission is the main route of transmission in vector-free areas. Congenital Chagas disease refers specifically to cases arising from this route of transmission. This work evaluates the clinical sensitivity of two qPCR techniques for diagnosis of congenital Chagas disease. The study was developed in the National Institute of parasitology (NIP), Argentina, and Pan-American Health Organization/ Word Health Organization Collaborating Center for Chagas Disease. Between July 2014 and May 2018, a prospective cohort study was carried out with 499 children born to seropositive for T. cruzi infection included. The performance of qPCR techniques was compared with the gold standard diagnostic algorithm for Congenital Chagas disease (CCD-GS), which comprises performing more than one parasitological test on children from birth until nine months of age, and serology from ten months of age. Of the 961 babies born to women seropositive for Chagas disease who were attended at the NIP laboratory, 462 did not meet the study inclusion criteria; 22 cases were diagnosed with congenital Chagas disease. qPCR showed 100% clinical sensitivity and 98 to 100% clinical specificity for the diagnosis of congenital Chagas disease compared with CCD-GS algorithm. The results obtained in this study demonstrate the clinical accuracy and effectiveness of qPCR SatDNA and qPCR kDNA for diagnosis of congenital Chagas disease. It could be a powerful tool for chagas test and treat strategies to reduce late complications of the disease. This work was financed by the INP Dr. Mario Fatala Chaben, ANLIS Dr. Carlos G. Malbran.

Microgeographical Variation in Dirofilaria immitis Prevalence in Dogs in Suburban and Urban Areas of Rio De Janeiro, Brazil.

Dirofilariosis, a mosquito-borne disease caused by Dirofilaria immitis and Dirofilaria repens, affects canids, felids and occasionally humans. Recent evidence suggests that D. immitis prevalence is rising in the canine populations in several areas of Brazil, even those historically considered to be non-endemic, highlighting the need for ongoing surveillance. However, prevalence studies are frequently based on inference from single diagnostic methods, and it is acknowledged that this may lead to biases and an underestimation of the disease situation. Thus, this cross-sectional study aimed to assess the prevalence of Dirofilaria spp. infection in dogs and cats across suburban and urban areas in Rio de Janeiro, Brazil. The investigation was conducted using a multi-diagnostic approach with binomial generalised linear models (GLMs) applied to investigate the association between the occurrence of infection and potential disease determinants. Blood samples from 497 dogs and 107 cats, collected at a veterinary mobile neutering unit, underwent parasitological, serological and molecular testing to detect Dirofilaria spp. infections. Among the dog samples, an overall 7.44% tested positive for D. immitis infection, while D. repens was not detected. In contrast, no cat samples tested positive for either parasite species. Prevalence was notably higher in suburban areas (14.47%) than in the urban area of Rio de Janeiro (6.17%) (p = 0.003). Dogs aged over two years had a higher risk of being infected (p < 0.001), while features such as breed, sex, size, coat type, use of macrocyclic lactones or access to outdoor spaces were not found to be predictive factors. This study reinforces the evidence that the prevalence of D. immitis is increasing in areas previously considered to have low prevalence or to be non-endemic. It also suggests that environmental and socioeconomic factors may play a key role in determining the current epidemiological status of this parasite. These findings highlight the need for increased monitoring in high-risk areas, targeted prevention and public awareness to address this emerging One Health concern.

Standardization and Evaluation of the LAMP Technique for the Diagnosis of Canine Visceral Leishmaniasis in Conjunctival Swab Samples Using DNA Extracted by a Silica Column and Boiling.

The diagnosis of canine visceral leishmaniasis (CVL) presents a challenge due to a variety of non-specific clinical signs. The available tests have low sensitivity. This study aimed to standardize and evaluate the loop-mediated isothermal amplification technique with K26 target (K26-LAMP) for diagnosis of CVL in conjunctival swab (CS) DNA samples extracted through a silica column commercial kit (SW-kit) and boiling (SW-DB) and to compare sensitivity with conventional PCR (kDNA-cPCR) and quantitative real-time PCR (18S-qPCR). Clinical samples of CSs were collected from 54 dogs after reactive serology tests. Positive parasitological and/or histological tests were used as inclusion criteria for a sensitivity analysis. A total of 79.2% (43/54) of dogs without clinical signs or with mild, moderate, or severe clinical signs were included in the study. The sensitivity results of K26-LAMP, kDNA-cPCR, and 18S-qPCR were 72.1%, 81.4%, and 80.5% with the SW-kit and 97.2%, 95.2%, and 57.1% with SW-DB, respectively. In all techniques, the proportion of positives was higher in the group with severe clinical disease, with statistically significant differences in the K26-LAMP and 18S-qPCR techniques being seen with the SW-kit. The results obtained with LAMP for CS samples are promising and its performance is similar to other techniques.

Chronic Strongyloides stercoralis infection in Fijian migrants to the UK.

Introduction. Strongyloides stercoralis, the human threadworm, is a parasitic nematode with global distribution, estimated to infect over 600 million people. Chronic infection is often asymptomatic, but hyperinfection and dissemination syndromes can occur in the immunosuppressed with high case fatality rates. Whilst strongyloidiasis is endemic in Fiji, its prevalence in Fijian migrant groups in the UK is unknown.Gap Statement. No previous studies have been conducted on the prevalence of Strongyloides and other gastrointestinal parasites (GIPs) in Fijian migrants to the UK.Aim. We conducted a cross-sectional study of the prevalence of GIPs in a Fijian migrant population.Methodology. Participants completed a questionnaire on residence, travel and clinical symptoms and were asked to provide a serum sample for S. stercoralis IgG ELISA, venous blood samples for eosinophil count and a faecal sample for charcoal culture, multiplex real-time PCR (rtPCR) and microscopy after formalin-ethyl acetate concentration. Sequencing was performed on pooled Strongyloides larvae for nuclear 18S rRNA hyper-variable regions (HVRs) I and IV.Results. A total of 250 participants (94% male) with median (range) age 37 (20-51) years entered the study, 15 (1-24) years since leaving Fiji. S. stercoralis IgG ELISA was positive in 87/248 (35.1 %) and 14/74 (18.9 %) had a GIP detected in faeces. This included 7/74 (9.5 %) with Strongyloides and 5/74 (6.8 %) with hookworms. Dermatological symptoms were more common in those with Strongyloides, and eosinophilia (>0.5×109 cells per litre) was present in 55.6% of those with positive S. stercoralis IgG. rtPCR was the most sensitive faecal diagnostic test for Strongyloides and hookworms in faeces. Sequences of nuclear 18S rRNA for HVRs I and IV confirmed the presence of S. stercoralis.Conclusion. This first cross-sectional study in Fijian migrants found a high rate of chronic infection with GIPs, particularly S. stercoralis. Faecal microscopy was insensitive compared to charcoal culture, rtPCR or serology, demonstrating the importance of specialist parasitological tests when investigating people with a suspected chronic infection. Our study highlights an overlooked burden of strongyloidiasis in the UK and has implications for screening and treatment programmes in Fiji and for migrants from Fiji.

First report of Dicrocoelium sp. In hoary fox (Lycalopex vetulus lund 1842) (carnivora: canidae) feces

The hoary fox (Pseudalopex vetulus), endemic to Brazil's Cerrado region, is a wild canid species facing ecological pressures from urbanization and habitat fragmentation. These pressures increase the risk of pathogen transmission between wild and domestic animals, particularly in rural and peri-urban areas. This study reports the first instance of Dicrocoelium sp. and Hepatozoon spp. infections in a wild female hoary fox in Southeast Brazil. The fox, presented with neurological disorders, dehydration, and poor nutritional condition, was hospitalized and subjected to a series of parasitological and clinical tests. Blood smears and fecal examinations revealed the presence of Hepatozoon sp. gametocytes and a significant number of Dicrocoelium eggs. The fox received treatment with albendazole, enrofloxacin, and N-acetylcysteine, showing progressive improvement before being reintroduced to its habitat. This study highlights the significance of parasitic infections in wild canids, emphasizing the need for further research on the ecological and health impacts of such infections. It also underscores the importance of comprehensive fecal and hematological examinations in the clinical assessment of wild fauna and raises awareness about the potential of wild canids as reservoirs for zoonotic parasites. KEY WORDS: Zoonosis; one health; canids.

Leishmaniasis in Tuscany: investigation of hidden human cases and comparison with canine incidence

Abstract The Mediterranean basin hosts endemic areas for leishmaniasis, mainly attributable to the presence of sand flies. Leishmaniasis has gained importance in Europe due to climate change, globalisation and migration. For at least the last 10 years in Tuscany, Italy, there has been an important discrepancy between official notifications of visceral and cutaneous leishmaniasis cases in humans and the number of hospital admissions, highlighting an important underestimation, despite mandatory notification. Moreover, official reports have increased alarmingly in the last three years. Human leishmaniasis does not always require hospitalisation: it often presents cutaneous manifestations, whereas in the immunocompetent individual it may not occur at all. For this reason, it is reasonable to assume that the number of cases is higher than the number of hospitalisations. The project aims to investigate the underreporting of human leishmaniasis cases from 2014 to 2023 by analysing laboratory diagnoses (considering any laboratory test that, according to the Italian Ministry of Health guidelines, is useful to identify a ‘confirmed case’, thus serological tests; parasitological tests; culture tests; PCR). To estimate the extent of underreporting, the capture-recapture method will be used, taking into account laboratory data, official reports and hospital data. Subsequently, a comparison will be made with the incidence, in the same territory, of veterinary cases, particularly in dogs, using data obtained from the Lazio-Tuscany Zooprophylactic Institute, to compare the trend of human and canine cases over time. Analyses are in progress and the results will be presented at the conference. Poor surveillance leads to ineffective prevention policies. The impact of the project is to strengthen the leishmaniasis surveillance system in Tuscany in order to reinforce prevention systems and face emergencies. Key messages • Due to climate change, globalization, and migration, there has been an increase in diagnoses of vector-borne diseases, including Leishmaniasis. • Leishmaniasis surveillance need to be strengthened in order to enhance prevention systems and address emergencies.

Concomitant Serological and Molecular Methods for Strongyloides stercoralis Screening in an Endemic Area of Spain.

Strongyloidiasis is a widespread parasitic disease that can be life-threatening in immunosuppressed people. In the Mediterranean basin, autochthonous cases coexist with imported ones. We aimed to assess the utility of different screening methods, along with the frequency of strongyloidiasis and its associated risk factors in migrants and the native population. This cross-sectional study took place from 2019 to 2022 in the area of the Vega Baja Hospital in Alicante, Spain. Screening was performed in people who were immunosuppressed, at risk of immunosuppression, with blood asymptomatic eosinophilia, and in asymptomatic people from highly endemic countries. Screening methods were serological techniques (ELISA), stool parasitological tests (fecal concentration methods and agar plate culture), and a stool molecular test (PCR). Of the 168 participants (62.5% males, 53.0% migrants, 36.3% immunosuppressed, median age 57 years), 14 (8.3%) had confirmed strongyloidiasis, where 6 were confirmed by serology, 4 by PCR, and 4 by both methods. Overall, 9% of the migrants and 7.6% of the native-born patients were infected. Elevated IgE and hemoglobin and Latin American origin were associated with strongyloidiasis diagnosis. Screening with serology alone would have missed 28.6% of cases. We conclude that strongyloidiasis prevalence is high in our population, both in native and migrant groups, and stool PCR is a useful tool to increase case detection.

Rapid and reliable detection of Leishmania antibodies in canine serum with double-antigen sandwich homogeneous chemical luminescence

BackgroundLeishmaniasis, caused by Leishmania spp. parasites, is an important zoonotic disease globally, posing severe threats to humans and animals. In the absence of effective vaccines, reliable serological diagnostic methods are critical for disease control. However, the enzyme-linked immunosorbent assay (ELISA) and immunochromatographic assay have limitations due to complexity, time required and/or sensitivity. Therefore, our objective was to develop an accurate, rapid and user-friendly detection method of canine leishmania antibody based on double-antigen sandwich homogeneous chemical luminescence.MethodsHomogeneous chemiluminescent technology was employed, and expressed recombinant fusion proteins containing full-length K9, K39 and K26 repeat sequences were used as diagnostic antigens. To establish a dual-antigen sandwich serological assay capable of detecting various antibody types, a factorial design was used to optimize concentrations of diagnostic antigen-receptor microspheres and of biotinylated diagnostic antigens, as well as of reaction solution composition and reaction duration. To evaluate and validate this newly developed method, we collected 41 Leishmania-positive serum samples, 30 Leishmania-negative control serum samples and 78 clinical serum samples for which no diagnostic information was available. Comparative analyses were performed using parasitological testing and an indirect ELISA as reference methods, focusing on diagnostic sensitivity and specificity.ResultsSodium dodecyl sulfate–polyacrylamide gel electrophoresis confirmed the purification of the diagnostic antigens, which exhibited clear bands without impurities. Based on results from the 41 Leishmania-positive samples and 30 Leishmania-negative samples, there was sufficient sensitivity to detect samples diluted up to 256-fold, with analytical specificity of 100%. Overall diagnostic sensitivity was 100% and diagnostic specificity was 93.3%. Diagnostic performance was highly consistent between the newly developed method and the indirect ELISA (Kappa = 0.82, P < 0.01). Testing could be completed within 35 min with the new methodConclusionsWe have developed a novel double-antigen sandwich homogeneous chemical luminescence method to detect canine Leishmania antibodies, with high sensitively and specificity, a short incubation interval and a simple protocol. This streamlined approach not only offers a sensitive and efficient method for clinical diagnosis but also has great potential for use in automated testing.Graphical

Prevalence of Helminths in Small Ruminant Farms and Evaluation of Control Practices Used to Counter Anthelmintic Resistance in Southern Italy.

Anthelmintic resistance in small ruminants is a serious worldwide problem. To reduce their spread, it is essential to know the prevalence of helminths on farms and the control practices adopted. As these studies in the Calabria region of southern Italy are fragmentary and outdated, a study on the prevalence of helminths in small ruminant holdings in this area has been conducted. The measures implemented to control helminths were also evaluated through questionnaires administered to farmers. In particular, on 90 farms (45 sheep and 45 goats), 1800 faecal samples from 900 sheep and 900 goats were collected in the spring. Using the FLOTAC dual technique, parasitological examinations demonstrated the presence of gastrointestinal nematodes in 100% of sheep and goat farms, followed by Nematodirus spp. (84.44% sheep and 48.89% goats), Moniezia spp. (73.33% sheep and 35.56% goats), Trichuris ovis (48.89% sheep and 42.22% goats), lungworms (28.89% sheep and 42.22% goats), Strongyloides papillosus (40% sheep and 26.67% goats), Dicrocoelium dendriticum (13.33% sheep and 26.67% goats), Calicophoron daubneyi (6.67% sheep and 31.11% goats), Fasciola hepatica (6.67% sheep and 4.44% goats), and Skrjabinema ovis (4.44% sheep and goats). The questionnaires showed that 82% and 85% of the farmers had applied pasture rotation, and that 93.3% and 86.6% had used anthelmintics in the previous year for sheep and goats, respectively. Only 24.4% of sheep farmers and 11.3% of goat farmers had carried out parasitological tests prior to treatments. The most used classes of anthelmintics were macrocyclic lactones and benzimidazoles, and only in 21.6% and 15.6%, for sheep and goats, respectively, was drug rotation carried out. These results denote that helminths represent a health problem for small ruminants and highlight a lack of knowledge of parasite control strategies among farmers. In these conditions, anthelmintic resistance phenomena could develop over time. Therefore, it is necessary to implement all possible strategies for the control of helminths, and to prevent the spread of anthelmintic resistance phenomena on farms in southern Italy.

Health workers’ adherence to malaria case management protocols in Northern Sudan: a qualitative study

BackgroundNonadherence to national standards for malaria diagnosis and treatment has been reported in Sudan. In this study, qualitative research examined the clinical domains of nonadherence, factors influencing nonadherent practices and health workers’ views on how to improve adherence.MethodsIn September 2023, five Focus Group Discussions (FGDs) were undertaken with 104 health workers from 42 health facilities in Sudan’s Northern State. The participants included medical assistants, doctors, nurses, laboratory personnel, pharmacists and public health officers. The FGDs followed a semi-structured guide reflecting the national malaria case management protocol. Qualitative thematic analysis was performed.ResultsNonadherent practices included disregarding parasitological test results, suboptimal paediatric artemether–lumefantrine (AL) dosing, lack of counselling, use of prohibited artemether injections for uncomplicated and severe malaria, artesunate dose approximations and suboptimal preparations, lack of AL follow on treatment for severe malaria; and rare use of primaquine for radical Plasmodium vivax treatment and dihydroartemisinin-piperaquine as the second-line treatment for uncomplicated malaria. Factors influencing nonadherence included stock-outs of anti-malarials and RDTs; staff shortages; lack of training, job aids and supervision; malpractice by specialists; distrust of malaria microscopy and RDTs; and patient pressure for diagnosis and treatment. Health workers recommended strengthening the supply chain; hiring personnel; providing in-service protocol training including specialists; establishing external quality assurance for malaria diagnosis; and providing onsite supportive supervision and public health campaigns.ConclusionsThis study revealed a broad spectrum of behavioural and systemic challenges in malaria management among frontline health workers in Northern Sudan, including nonadherence to protocols due to resource shortages, training gaps, a lack of supportive supervision and patient pressure. These insights, including health workers’ views about improvements, will inform evidence-based interventions by Sudan’s National Malaria Control Programme to improve health systems readiness and the quality of malaria case management.

Gastrointestinal Parasites in Owned Dogs in Serbia: Prevalence and Risk Factors.

Dogs are the most popular pets worldwide. Close contact between dogs and people increases the risk of transmission of various zoonotic parasitic infections. Given the importance of veterinary medicine in preserving the One Health concept, the aim of this research was to identify intestinal parasites that may have zoonotic potential and to evaluate risk factors (individual and environmental). The research was conducted in Serbia in 2022 and 2023 on 382 owned dogs, using qualitative methods of coprological examination with a concentration on parasitic elements. The overall prevalence of intestinal parasites was 62.6%, with the following detected: protozoa: Cystoisospora spp. (9.2%), Sarcocystis spp. (4.5%), Neospora caninum/Hammondia spp. (3.7%), Giardia intestinalis (11.8%); nematoda: Toxocara canis (11.5%), Toxascaris leonina (4.2%), family Ancylostomatidae (38.0%), Trichuris vulpis (21.5%), Capillaria spp. (10.5%); trematoda: Alaria alata (1.6%) and cestodes from the Taeniidae family (1.3%). Factors like age, size and coat length, as well as the way of living, attitude and diet were linked to a significantly higher (p < 0.05) prevalence of intestinal parasites. Based on the results of coprological diagnostics, this research indicates the importance of educating dog owners, conducting routine parasitological tests on their pets and regular deworming strategies.

Epidemiological analysis of Dirofilaria immitis (Spirurida: Onchocercidae) infecting pet dogs (Canis lupus familiaris, Linnaeus, 1758) in Baixada Fluminense, Rio de Janeiro.

Dirofilaria immitis infection is routinely detected in dogs during veterinary care in tropical and subtropical regions worldwide. Parasitological tests for the detection of this infection are routinely performed only in areas with a high prevalence. Baixada Fluminense, a region in Rio de Janeiro, was considered heartworm-free until local veterinarians began to receive blood exams results indicating the presence of microfilariae (MF). A laboratory database was hence used to collect data from 2017 to 2020 to understand the extent of spread of the parasite in this area. The results of complete blood count analysis and MF or heartworm antigen detection tests conducted on canine samples sent from veterinary clinics in Baixada Fluminense (Magé, Duque de Caxias, Guapimirim, Nova Iguaçu, and São João de Meriti municipalities) were included. In total, the results of 16,314 hematological tests were considered. The overall prevalence of D. immitis was 3.4% (554/16,314), considering that only one test result was obtained per animal on the same day. This study is highly relevant because it indicates the spreading geographic distribution of the worms, heightens awareness among local health professionals and the general population, and encourages compliance with prophylactic measures to prevent further spread of parasite.

Динамика на някои биохимични показатели при две местни породи овце в зависимост от сезона, надморската височина и наличието на паразити

The aim of the present study is to evaluate the values of some enzymes and metabolites - ALT, AST, ALP, serum iron and glucose of two herds of Bulgarian native sheep breeds raised at different altitudes, depending on the season, their body condition score (BCS) and the presence of gastrointestinal parasites. The research was conducted with 20 animals each in farms in the village of Mirkovo (715 m above sea level) with Sofia (Elinpelin) sheep breed and in the village of Gubesh (1040 m above sea level) with Central Balkan sheep breed. Body condition assessment was performed during sampling by one individual using an adapted system. Fresh faeces (total sample) were also collected for parasitological tests (coproovoscopy and coprolarvoscopy). Analysis of biochemical parameters was performed in extracted blood plasma with a BTS-350 semi-automatic biochemical analyzer. AST and glucose levels were normal, ALP and Fe were lower in both seasons, and ALT was higher than reference values limits only in summer for both breeds. There were significant differences between the two breeds in summer for BCS, ALT and glucose (p&lt;0.001, p&lt;0.05 and p&lt;0.01 respectively), in winter - only for Fe (p&lt;0.001) and in both seasons for HCT (p&lt;0.001 and p&lt;0.01 respectively), AST (p&lt;0.001 and p&lt;0.05 respectively) and ALP (p&lt;0.001 each). Regarding the activity of AST, there are no significant differences between the two seasons and in both breeds, however, there are such differences only in Central Balkan sheep for BCS and Fe (respectively p&lt;0.01 and p&lt;0.001), and in both breeds together with different degrees of freedom - for all other indicators (HCT, ALT, ALP and glucose).

Leucemia linfocítica crônica em cão Shih-Tzu

Chronic Lymphocytic Leukemia (CLL) is defined as a malignant neoplasm originating from hematopoietic stem cells in the bone marrow, where the lymphocytes found in circulation are mature and well-differentiated. The disease is rare in dogs and cats and, when it occurs, it typically affects elderly animals. This study presents an atypical clinical case of a two-years-old Shih Tzu diagnosed with chronic lymphocytic leukemia. The clinical state of the patient included characteristic signs of CLL, such as lymphocytosis, lymphadenomegaly, splenomegaly, and hepatomegaly. Observed symptoms included lethargy, fatigue, panting and enlargement of the mandibular lymph nodes. Additionally, anemia and thrombocytopenia were identified, in alignment with common patterns in dogs with CLL. The diagnosis was achieved through various examinations, including ultrasound, radiography, complete blood count, serum biochemistry, cytology and parasitological tests (4DX and leishmaniasis). However, definitive confirmation of CLL was obtained through bone marrow examination. In response to the diagnosis, therapeutic intervention followed the protocol established in the literature, involving the administration of Chlorambucil and Prednisone. The results of the applied treatments reflected positively on the patient, demonstrating clinical improvement and stability. The adopted therapeutic protocol aimed to control symptoms and prolong the life of the affected animal. The objective of this study is to contribute to epidemiological data, elucidate the pathogenic course and seek more effective diagnostic and therapeutic approaches for CLL in dogs. The detailed investigation of this case and the dissemination of results aim to provide a solid foundation for future research, with the goal of improving the quality of life for dogs affected by chronic lymphocytic leukemia.

Aetiology of non-malaria acute febrile illness fever in children in rural Guinea-Bissau: a prospective cross-sectional investigation.

With growing use of parasitological tests to detect malaria and decreasing incidence of the disease in Africa; it becomes necessary to increase the understanding of causes of non-malaria acute febrile illness (NMAFI) towards providing appropriate case management. This research investigates causes of NMAFI in pediatric out-patients in rural Guinea-Bissau. Children 0-5 years presenting acute fever (≥38°) or history of fever, negative malaria rapid diagnostic test (mRDT) and no signs of specific disease were recruited at the out-patient clinic of 3 health facilities in Bafatá province during 54 consecutive weeks (dry and rainy season). Medical history was recorded and blood, nasopharyngeal, stool and urine samples were collected and tested for the presence of 38 different potential aetiological causes of fever. Samples from 741 children were analysed, the protocol was successful in determining a probable aetiological cause of acute fever in 544 (73.61%) cases. Respiratory viruses were the most frequently identified pathogens, present in the nasopharynx samples of 435 (58.86%) cases, followed by bacteria detected in 167 (22.60%) samples. Despite presenting negative mRDTs, P. falciparum was identified in samples of 24 (3.25%) patients. This research provides a description of the aetiological causes of NMAFI in West African context. Evidence of viral infections were more commonly found than bacteria or parasites.

An α-Gal antigenic surrogate as a biomarker of treatment evaluation in Trypanosoma cruzi-infected children. A retrospective cohort study.

Proper evaluation of therapeutic responses in Chagas disease is hampered by the prolonged persistence of antibodies to Trypanosoma cruzi measured by conventional serological tests and by the lack of sensitivity of parasitological tests. Previous studies indicated that tGPI-mucins, an α-Gal (α-d-Galp(1→3)-β-d-Galp(1→4)-d-GlcNAc)-rich fraction obtained from T. cruzi trypomastigotes surface coat, elicit a strong and protective antibody response in infected individuals, which disappears soon after successful treatment. The cost and technical difficulties associated with tGPI-mucins preparation, however, preclude its routine implementation in clinical settings. We herein developed a neoglycoprotein consisting of a BSA scaffold decorated with several units of a synthetic α-Gal antigenic surrogate (α-d-Galp(1→3)-β-d-Galp(1→4)-β-d-Glcp). Serological responses to this reagent, termed NGP-Tri, were monitored by means of an in-house enzyme-linked immunosorbent assay (α-Gal-ELISA) in a cohort of 82 T. cruzi-infected and Benznidazole- or Nifurtimox-treated children (3 days to 16 years-old). This cohort was split into 3 groups based on the age of patients at the time of treatment initiation: Group 1 comprised 24 babies (3 days to 5 months-old; median = 26 days-old), Group 2 comprised 31 children (7 months to 3 years-old; median = 1.0-year-old) and Group 3 comprised 26 patients (3 to 16 years-old; median = 8.4 years-old). A second, control cohort (Group 4) included 39 non-infected infants (3 days to 5 months-old; median = 31 days-old) born to T. cruzi-infected mothers. Despite its suboptimal seroprevalence (58.4%), α-Gal-ELISA yielded shorter median time values of negativization (23 months [IC 95% 7 to 36 months] vs 60 months [IC 95% 15 to 83 months]; p = 0.0016) and higher rate of patient negative seroconversion (89.2% vs 43.2%, p < 0.005) as compared to conventional serological methods. The same effect was verified for every Group, when analyzed separately. Most remarkably, 14 out of 24 (58.3%) patients from Group 3 achieved negative seroconversion for α-Gal-ELISA while none of them were able to negativize for conventional serology. Detailed analysis of patients showing unconventional serological responses suggested that, in addition to providing a novel tool to shorten follow-up periods after chemotherapy, the α-Gal-ELISA may assist in other diagnostic needs in pediatric Chagas disease. The tools evaluated here provide the cornerstone for the development of an efficacious, reliable, and straightforward post-therapeutic marker for pediatric Chagas disease.

Evaluation of Malaria Microscopy Diagnostic Performance at 40 Public Health Facilities in Abidjan, Côte d'Ivoire in 2020.

Microscopic diagnosis of Giemsa-stained thick and thin blood films remained the gold standard laboratory method for the diagnosis of malaria. In this context, we felt it was important to conduct this evaluation with 40 public medical biology laboratories (MBLs) in the Abidjan 1 health region that perform blood parasitology tests to improve their implementation process. This descriptive and analytical study took place in July 2020 and involved participating laboratories (PLs) from the public sector in Abidjan. A set of 3 blood smear slides of variable parasite densities (PDs) with assigned values (AVs) of parasite densities and assigned Plasmodium species was used. The criterion for establishing the parasite density compliance interval was assigned values of ± 25%, and the performance rates were compared to the 80% recommended by the WHO for the African region. Nearly a quarter (11/40) of the participating laboratories had a compliance rate greater than 80%, including 10 with a performance of 100% for the ability to identify parasites. Regarding identifying plasmodial species, a concordance rate of 100% was obtained for slide 1 for Plasmodium falciparum, while this rate was 20% for slide 2 for Plasmodium ovale. For parasite densities < 200/µl, 87.5% of the participating laboratories (PLs) had a performance rate lower than 80%, while 95% of these PLs had a performance rate higher than 80% for parasitaemia > 2000/µl. There is a need to strengthen adapted to low parasitaemia, to improve the biological confirmation of malaria in Côte d'Ivoire.