Frecuencia serológica y molecular de Leishmania spp y Trypanosoma cruzi y factores asociados en perros con signos sugestivos de infección por tripanosomátidos en Quintana Roo, México

A cross-sectional study was conducted with a sample of 40 dogs with clinical signs suggestive of infection by Leishmania spp and Trypanosoma cruzi in the municipality of Lázaro Cárdenas, Quintana Roo, Mexico. Blood, skin and bone marrow samples were taken. Indirect immunofluorescence (IFI) tests for detection of anti-Leishmania infantum and immunochromatic test for the detection of anti-Trypanosoma cruzi antibodies in serum were performed. In addition, molecular analyses were performed using PCR to detect Leishmania spp in bone marrow and skin and T. cruzi in bone marrow. Histopathological analyses of the skin and cytology of the bone marrow were carried out in search of lesions and trypanosomatid agents. The results showed that 3 (7.5%) and 6 (15%) dogs with anti-L. infantum and T. cruzi antibodies were detected, respectively. Molecular analyses detected 17 dogs positive for infection by T. cruzi (42.5%) and none for infection by Leishmania spp. Dogs with low body condition showed greater seropositivity to T. cruzi using Fisher’s exact test (p<0.05). Parasites of the genus Leishmania did not actively participate in the pathological processes of the animals; however, a high frequency of animals infected with T. cruzi was observed.Se realizó un estudio transversal con una muestra de 40 perros con signos clínicos sugestivos de infección por Leishmania spp y Trypanosoma cruzi en el municipio de Lázaro Cárdenas, Quintana Roo, México. Se tomaron muestras de sangre, piel y médula ósea. Se realizaron las pruebas de inmunofluorescencia indirecta (IFI) para detección de anticuerpos anti-Leishmania infantum e inmunocromática para detección de anticuerpos anti-Trypanosoma cruzi en suero. Además, se realizaron análisis moleculares, mediante PCR para la detección de Leishmania spp en médula ósea y piel y T. cruzi en médula ósea. Se realizaron análisis histopatológicos de piel y citológicos de médula ósea en busca de lesiones y agentes tripanosomátidos. Se detectaron 3 (7.5%) y 6 (15%) perros con anticuerpos anti-L. infantum y T. cruzi, respectivamente. En los análisis moleculares se detectaron 17 perros positivos a la infección por T. cruzi (42.5%) y ninguno a la infección por Leishmania spp. Los perros con baja condición corporal presentaron mayor seropositividad a T. cruzi mediante la prueba exacta de Fisher (p<0.05). Los parásitos del género Leishmania no participaron activamente en los procesos patológicos de los animales estudiados; sin embargo, se observa una alta frecuencia de animales infectados por T. cruzi

Casos clínicos virtuales para el aprendizaje de las Enfermedades Parasitarias en Veterinaria

Depto. de Sanidad AnimalFac. de VeterinariaFALSEUCMsubmitte

Hemolysis, icterus and lipemia interfere with the determination of two oxidative stress biomarkers in canine serum

2023 Acuerdos transformativos CRUEBackground: Oxidative stress has been proven to play a role in numerous human and canine diseases. Among the biomarkers of oxidative stress, Thiobarbituric Acid Reactive Substances (TBARS) and Total Antioxidant Status (TAS) are two of the most widely used. Preanalytical factors are crucial for obtaining accurate results in these assays. Hemolysis, icterus and lipemia (HIL) are common sources of preanalytical errors in the laboratory; however, limited information is available regarding the considerations for canine specimens. Therefore, the objective of this study was to evaluate the potential interferences of HIL in the determination of TBARS and TAS in canine serum. Methods: Solutions of pooled canine serum samples were prepared by adding increasing concentrations of hemolysate, bilirubin and a synthetic lipid emulsion. TBARS and TAS were determined, and biases from the control value caused by the interfering substances were calculated. Results: Hemolysis, icterus and lipemia induced significant interferences on TBARS and TAS, albeit to varying degrees depending on the specific biomarker and interfering substance. TBARS appeared to be more susceptible to interferences in this study. Slight hemolysis, moderate icterus and slight lipemia caused notable deviations in TBARS values, surpassing the acceptable threshold for interference. TAS assay was also affected by HIL, although to a lesser extent compared to TBARS. Significant biases from TAS control value were observed when icterus was moderate, and when hemolysis and lipemia were more pronounced. Conclusions: In light of our results, we conclude that hemolyzed, icteric and lipemic specimens are not suitable for TBARS and TAS determination in canine serum. Our findings hold considerable practical utility, as a simple visual inspection would be sufficient for identifying and excluding such specimens.Depto. de Medicina y Cirugía AnimalDepto. de Sanidad AnimalFac. de VeterinariaCentro de Vigilancia Sanitaria Veterinaria (VISAVET)TRUEpubAPC financiada por la UC

Cystoisospora spp. infection at a dog breeding facility in the Madrid region: Infection rate and clinical management based on toltrazuril metaphylaxis

Canine coccidiosis caused by Cystoisospora canis and Cystoisospora ohioensis-complex is common in kennels. While often underestimated, coccidiosis may cause severe clinical signs in puppies and sometimes even lead to death, so preventative measures are important. This study examines Cystoisospora spp. infection at a Labrador retriever breeding facility in Madrid, Spain. To identify environmental factors associated with infection, dams were examined throughout a reproductive cycle (from oestrus to 60 days postpartum) and their puppies during their first 60 days of life. Also assessed was the efficacy of combined treatment with emodepside (0.9 mg/ml) and toltrazuril (18 mg/ml) at a dose of 0.5 ml/kg of weight, equivalent to 0.45 mg/kg and 9 mg/kg, respectively, in puppies on day 35 of life. Oocyst shedding was detected in 4.6–18.6% of 45 dams examined and in 2.2–9.1% of their litters (315 puppies). In both cases, peak opg elimination was recorded on day 30 postpartum/of life. The species of Cystoisospora detected were C. canis (91.3%) and C. ohioensis-complex (8.7%). While in both dams and puppies opg counts were higher in autumn when rainfall was at its highest, correlation between opg and rainfall emerged as significant only in puppies (p = 0.031). The treatment of 35 day-old puppies with toltrazuril was 100% effective in controlling this infection in the kennel. Our findings therefore suggest the need for a strict hygiene regime and the use of toltrazuril as blanket treatment to reduce Cystoisospora transmission in dog breeding facilities.Depto. de Sanidad AnimalFac. de VeterinariaTRUEpu

A blinded, randomized and controlled multicenter clinical trial to assess the efficacy and safety of Leisguard® as an immunotherapeutic treatment for healthy Leishmania infantum-seropositive dogs

2023 Acuerdos transformativos CRUEBackground: Domperidone (Leisguard®) is an immunomodulatory drug used as a preventive measure in healthy dogs. However, no studies have been published in healthy Leishmania infantum-seropositive dogs. The aim of this study was to evaluate the clinical efficacy and safety of domperidone as immunotherapy in Leishmania-seropositive healthy dogs. Methods: Sixty-seven dogs were treated with domperidone at 0.5 mg/kg and 44 dogs received placebo, once daily for 4 consecutive weeks. Monthly treatments were repeated every 4 months until the end of the 1-year follow-up period. Veterinary examinations were performed on days 0, 30, 120, 150, 240, 270 and 360. Samples of blood and urine were collected on days 0, 120, 240 and 360 for routine laboratory tests and quantitative in-house ELISA for the detection of L. infantum-specific antibodies. Furthermore, Leishmania real-time PCR and IFN-γ ELISA were performed at day 0 and the end of the study. Dogs that developed disease were withdrawn from the study and classified as sick dogs. Adverse drug reactions were reported. Results: Thirty dogs developed disease during the follow-up period: 13/67 (19.4%) in the group treated with domperidone and 17/44 (38.6%) in the placebo-treated group (P = 0.03). Low-seropositive dogs treated with domperidone (4/40, 9.1%) were significantly less likely to develop disease compared to low-seropositive dogs treated with placebo (7/24, 29.2%; P = 0.04), while no differences were found between domperidone (9/23, 39.1%) and placebo (10/20, 50%) in medium- to high-seropositive dogs. At the end of the study, a higher proportion of Leishmania PCR-positive dogs was observed in the placebo-treated group (16/33, 48.5%) compared to the domperidone group (13/51, 25.5%; P = 0.04). Furthermore, low-seropositive dogs treated with domperidone with an increase of IFN-γ concentration presented a higher increase than those treated with placebo at the end of the study. Four dogs treated with domperidone presented self-limiting diarrhea. Conclusions: Healthy dogs with low L. infantum antibody levels treated with domperidone were less likely to develop disease compared to placebo-treated dogs. Furthermore, domperidone presented a good safety profile.Depto. de Sanidad AnimalFac. de VeterinariaTRUEpubAPC financiada por la UC

Clinical and hematological follow-Up of long-term oral therapy with type-I interferon in cats naturally infected with feline Leukemia virus or felineiImmunodeficiency Virus

Feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV), two of the most important pathogens of cats, produce chronic systemic diseases with progressive death of cells involved in the immune response, ultimately leading to death. Immunostimulants is one of the few alternatives to the symptomatic treatment. In this study, 27 naturally FeLV-infected (FeLV+) and 31 naturally FIV-infected (FIV+) cats were administered orally by their owners 60 IU/day of recombinant human interferon alpha (rHuIFN-α) for four months in alternate weeks. Clinical status was evaluated and blood samples collected at four different visits or months (M): pretreatment (M0), mid-treatment (M2), end of treatment (M4), and 4–8 months after end of treatment (M10). Most cats ostensibly improved their clinical status, and many became asymptomatic. rHuIFN-α treatment improved the anemic processes observed at M0 (at least in cats with mild or moderate anemia) and leukocyte counts, including a more favorable CD4+/CD8+ ratio. An increase in the serum gammaglobulin concentration was seen in 80% of the cats. Despite observing an obvious favorable progress in the clinical, biopathological, and CD4+/CD8+ values during treatment, almost invariably all the parameters analyzed worsened after treatment discontinuation (M10), which suggests that the interferon-α protocol should be either extended or include additional cycles for a long-lasting benefit in FeLV+ and FIV+ cats

Tritrichomonas foetus infection in cats with diarrhea from denselyhoused origins

Tritrichomonas foetus is a protozoan parasite that has been recently identified as a causative agent of chronic diarrhea in domestic cats. Transmission of infection occurs by the fecal-oral route through direct contact among animals. Consequently, feline trichomonosis (FT) is more likely to be present in multi-cat environments. The objective of this work was to study the presence of T. foetus and some associated risk factors in cats from densely housed origins and with a reported history of chronic diarrhea. Animals enrolled in this study were family cats (n = 15) acquired from pet shops, shelters or breeding centers and cattery cats belonging to one breeding center (n = 28) and two cat shelters (A and B, n = 25 each). In the catteries, a follow-up analysis for a period of up to 2 months was also performed to determine the parasite shedding pattern in feces and the incidence of infection. Fecal samples were analyzed using in vitro culture and a PCR technique. T. foetus was detected in a total of 38.7% (36/93) of the cats with chronic diarrhea. Parasite infection was similarly detected in family cats and cattery animals (40% versus 38.4%). In the catteries, the parasite was detected in 50%, 44% and 20% of the animals from the breeding center and shelters A and B, respectively. The follow-up analysis showed that 58.3% of infected cats intermittently shed trophozoites in their feces, with an incidence of 23.1%. Investigation of potential risk factors showed that cats ≤1 year old were more likely to be infected than older cats (57.1% versus 27.3%; P < 0.05). No significant differences were found when sex and breed factors were studied. These results confirm the importance of FT as a cause of chronic diarrhea in cats and highlight the relevance of close contact conditions for T. foetus transmission.Depto. de Sanidad AnimalCAI Ciencias de la Tierra y ArqueometríaTRUEpu

First identification of Strongyloides stercoralis infection in a pet dog in Argentina, using integrated diagnostic approaches

2023 Acuerdos transformativos CRUEBackground: Strongyloides stercoralis is a soil-transmitted intestinal nematode with a complex life cycle that primarily affects humans, non-human primates, dogs, and occasionally cats. This study presents, to the best of our knowledge, the first case of S. stercoralis infection and its genotyping in a domestic dog from Argentina. Methods: The patient was a female wired-haired Teckel dog exhibiting recurrent coughing. Coproparasitological analysis using the Baermann technique revealed the presence of rhabditiform larvae morphologically compatible with S. stercoralis. To confirm this finding, molecular diagnosis (18S ribosomal RNA) and analysis of the cox1 gene were performed. Results: We identified a haplotype (HP20) that has previously only been related to S. stercoralis infection in dogs, but was found in the present study to be highly related to the haplotype (HP16) of a zoonotic variant and divergent from those previously described from human patients in Argentina. Furthermore, unlike in human cases following treatment with ivermectin, the dog was negative after moxidectin treatment according to polymerase chain reaction of the sampled faeces. Conclusions: This case report shows the importance of further investigation into potential transmission events and prevalences of S. stercoralis in dogs and humans in South America. The results reported here should also encourage future work that examines different scenarios of infection with S. stercoralis in dogs and humans with the aim of integrating clinical management, diagnosis, treatment and follow-up strategies in the quest for new approaches for the treatment of this disease in animals and humans. The findings support the adoption of a One Health approach, which recognizes the interconnectedness between animal and human health, in addressing parasitic infections such as strongyloidiasis.Depto. de Sanidad AnimalFac. de VeterinariaTRUEpubAPC financiada por la UC

Clinical validation of circulating immune complexes for use as a diagnostic marker of canine leishmaniosis

Author contributions: JS: Data curation, Formal analysis, Investigation, Methodology, Project administration, Resources, Writing – original draft, Writing – review & editing. NP: Investigation, Validation, Writing – review & editing. AM: Resources, Writing – review & editing. CC-L: Investigation, Validation, Writing – review & editing. JB: Resources, Writing – review & editing. RC: Resources, Writing – review & editing. MD: Resources, Writing – review & editing. JC: Conceptualization, Funding acquisition, Project administration, Supervision, Writing – review & editing. GM: Conceptualization, Funding acquisition, Methodology, Project administration, Resources, Supervision, Writing – review & editing.Canine leishmaniosis (CanL) is a systemic disease that affects dogs. When multiplication of the parasite cannot be controlled, dogs consistently show high levels of antigen and IgG antibodies, which lead to the formation of circulating immune complexes (CIC). Timely intervention to reduce the parasite load and CIC levels is crucial for preventing irreversible organ damage. However, a diagnostic test to quantify CIC levels is currently lacking. In this real-world study, we aimed to examine the performance of a new ELISA to measure CIC levels in dogs naturally infected with Leishmania infantum. Thirty-four dogs were treated according to their clinical condition and followed for 360 days. Before (day 0) and after treatment (days 30, 90, 180, 270, and 360), all dogs underwent a physical examination, and blood samples were obtained for CBC, biochemical profile, serum protein electrophoresis and IFAT. Serum PEG-precipitated CIC were determined by ELISA. Our results indicate higher CIC levels in dogs in advanced disease stages showing higher antibody titres (p  <  0.0001, r  =  0.735), anemia (p  <  0.0001), dysproteinemia (p  <  0.0001), and proteinuria (p  =  0.004). Importantly, dogs responding well to treatment exhibited declining CIC levels (p  <  0.0001), while in poor responders and those experiencing relapses, CIC were consistently elevated. CIC emerged as a robust discriminator of relapse, with an area under the curve (AUC) of 0.808. The optimal cut-off to accurately identify relapse was an optical density of 1.539. Our findings suggest that declining CIC levels should be expected in dogs showing a favorable treatment response. Conversely, in dogs displaying a poor response and recurrent clinical relapses, CIC levels will be high, emphasizing the need for vigilant monitoring. These findings suggest that CIC could serve as a valuable biomarker for disease progression, treatment efficacy, and relapse detection in CanL. Our study contributes to enhancing diagnostic approaches for CanL and underscores the potential of CIC as a complementary tool in veterinary practice. As we move forward, larger studies will be essential to confirm these findings and establish definitive cut-offs for clinical application.Depto. de Sanidad AnimalFac. de VeterinariaTRUEpu

LeishVet guidelines for the practical management of canine leishmaniosis

The LeishVet group has formed recommendations designed primarily to help the veterinary clinician in the management of canine leishmaniosis. The complexity of this zoonotic infection and the wide range of its clinical manifestations, from inapparent infection to severe disease, make the management of canine leishmaniosis challenging. The recommendations were constructed by combining a comprehensive review of evidence-based studies, extensive clinical experience and critical consensus opinion discussions. The guidelines presented here in a short version with graphical topic displays suggest standardized and rational approaches to the diagnosis, treatment, follow-up, control and prevention of canine leishmaniosis. A staging system that divides the disease into four stages is aimed at assisting the clinician in determining the appropriate therapy, forecasting prognosis, and implementing follow-up steps required for the management of the leishmaniosis patient