Failure of Miltefosine Treatment in Two Dogs with Natural Leishmania infantum Infection

Two dogs, with naturally acquired canine leishmaniasis, were treated orally with miltefosine (2 mg/kg q 24 hr) and allopurinol (10 mg/kg q 12 hr) for 28 days. Both dogs showed good initial response to therapy, with reduction in clinical signs and improvement of clinicopathological changes. However, in both dogs, clinical and clinicopathological abnormalities recurred 150 days after initial treatment and a second course of miltefosine and allopurinol was administered. One dog failed to respond to the 2nd cycle of miltefosine treatment and the other dog responded initially but suffered an early relapse. Treatment with meglumine antimoniate (100 mg/kg q 24 hr for a minimum of 4 weeks) was then started in both dogs. Both dogs showed rapid clinical and clinicopathological improvement and to date they have not received further treatment for 420 and 270 days, respectively. In view of the low number of antileishmanial drugs available and the fact that some of these are used in human as well as veterinary medicine, it is of paramount importance that drug resistance is monitored and documented

Ammonia concentration and bacterial evaluation of feline whole blood and packed red blood cell units stored for transfusion

Ammonia concentrations increase in human, canine and equine WB and PRBC units during storage. The aim of this study was to determine the effect of storage on ammonia concentration in feline WB and PRBC units stored in a veterinary blood bank and to evaluate possible correlations with bacterial contamination. Ammonia concentration was evaluated in 15 WB units and 2 PRBC units on day 1 and at the end of storage after 35 and 42 days, respectively. In an additional 5 WB units and 4 PRBC units ammonia concentrations were determined daily until the day the normal reference range was exceeded and then weekly to the end of storage. All units were evaluated for bacterial contamination. Ammonia increased markedly during storage as a linear function over time. On the 35th and 42th day of storage at 4±2°C mean±SD ammonia concentration reached 909±158 µg/dl and 1058±212 µg/dl in WB and PRBC units, respectively. Bacterial culture was negative in all units. High ammonia concentrations in stored WB and PRBC units could result in toxicity, particularly in feline recipients with liver failure, portosystemic shunts or those receiving large transfusion volumes. Clinical in vivo studies evaluating the effects on recipients should be performed

FELINE LEISHMANIASIS: SEROLOGICAL AND MOLECULAR DETECTION OF AN EMERGENT DISEASE IN A NON-ENDEMIC AREA OF NORTHERN ITALY

In recent decades feline leishmaniosis (FeL) has become an emerging disease, also in non-endemic areas for the canine infection. This study updates the epidemiological status for FeL in cats in northern Italy and compares results with previous studies of the same feline population. Co-infections with feline retroviruses FIV and Field were also investigated. Stray, shelter and owned cats from different cities in the Lombardy region of northern Italy, were prospectively randomly sampled between January 2020 and May 2021. A total of 255 cats were tested for L. infantum: 240/255 for antibodies by IFAT and 234/255 and 198/255 for Leishmania DNA by PCR on whole blood and lymph nodes, respectively. Rapid ELISA test was used to detect FIV or FeLV infection. Overall, 26/255 (10.2%) cats tested positive for L. infantum: in 8/26 cats Leishmania DNA was found in popliteal lymph nodes (Leishmania/ml range from 15 to 60), 6/26 were PCR positive on whole blood (Leishmania/ml range from 5 to 80) and 15/26 IFAT seropositive at titers ranging from 1:80 to 1:320. Two Leishmania infected cats were also FIV+FeLV coinfected, another was FIV positive and one was FeLV positive. A high prevalence of FeL was found in a non-endemic area of northern Italy, with an increasing trend in infection rates

DNA Metabarcoding and Isolation by Baiting Complement Each Other in Revealing Phytophthora Diversity in Anthropized and Natural Ecosystems

Isolation techniques supplemented by sequencing of DNA from axenic cultures have provided a robust methodology for the study of Phytophthora communities in agricultural and natural ecosystems. Recently, metabarcoding approaches have emerged as new paradigms for the detection of Phytophthora species in environmental samples. In this study, Illumina DNA metabarcoding and a conventional leaf baiting isolation technique were compared to unravel the variability of Phytophthora communities in different environments. Overall, 39 rhizosphere soil samples from a natural, a semi-natural and a horticultural small-scale ecosystem, respectively, were processed by both baiting and metabarcoding. Using both detection techniques, 28 out of 39 samples tested positive for Phytophthora. Overall, 1,406,613 Phytophthora internal transcribed spacer 1 (ITS1) sequences and 155 Phytophthora isolates were obtained, which grouped into 21 taxa, five retrieved exclusively by baiting (P. bilorbang; P. cryptogea; P. gonapodyides; P. parvispora and P. pseudocryptogea), 12 exclusively by metabarcoding (P. asparagi; P. occultans; P. psycrophila; P. syringae; P. aleatoria/P. cactorum; P. castanetorum/P. quercina; P. iranica-like; P. unknown sp. 1; P. unknown sp. 2; P. unknown sp. 3; P. unknown sp. 4; P. unknown sp. 5) and four with both techniques (P. citrophthora, P. multivora, P. nicotianae and P. plurivora). Both techniques complemented each other in describing the variability of Phytophthora communities from natural and managed ecosystems and revealing the presence of rare or undescribed Phytophthora taxa

An expert consensus on the recommendations for the use of biomarkers in Fabry disease

Fabry disease is an X-linked lysosomal storage disorder caused by the accumulation of glycosphingolipids in various tissues and body fluids, leading to progressive organ damage and life-threatening complications. Phenotypic classification is based on disease progression and severity and can be used to predict outcomes. Patients with a classic Fabry phenotype have little to no residual α-Gal A activity and have widespread organ involvement, whereas patients with a later-onset phenotype have residual α-Gal A activity and disease progression can be limited to a single organ, often the heart. Diagnosis and monitoring of patients with Fabry disease should therefore be individualized, and biomarkers are available to support with this. Disease-specific biomarkers are useful in the diagnosis of Fabry disease; non-disease-specific biomarkers may be useful to assess organ damage. For most biomarkers it can be challenging to prove they translate to differences in the risk of clinical events associated with Fabry disease. Therefore, careful monitoring of treatment outcomes and collection of prospective data in patients are needed. As we deepen our understanding of Fabry disease, it is important to regularly re-evaluate and appraise published evidence relating to biomarkers. In this article, we present the results of a literature review of evidence published between February 2017 and July 2020 on the impact of disease-specific treatment on biomarkers and provide an expert consensus on clinical recommendations for the use of those biomarkers.publishedVersio

Active turnover of genomic methylcytosine in pluripotent cells

Epigenetic plasticity underpins cell potency, but the extent to which active turnover of DNA methylation contributes to such plasticity is not known and the underlying pathways are poorly understood. Here we use metabolic labelling with stable isotopes and mass spectrometry to quantitatively address the global turnover of genomic methylcytidine (mdC), hydroxymethylcytidine (hmdC) and formylcytidine (fdC) across mouse pluripotent cell states. High rates of mdC/hmdC oxidation and fdC turnover characterize a formative-like pluripotent state. In primed pluripotent cells the global mdC turnover rate is about 3-6% faster than can be explained by passive dilution through DNA synthesis. While this active component is largely dependent on Tet-mediated mdC oxidation, we unveil additional oxidation-independent mdC turnover, possibly through DNA repair. This process accelerates upon acquisition of primed pluripotency and returns to low levels in lineage committed cells. Thus, in pluripotent cells active mdC turnover involves both mdC oxidation-dependent and independent processes

Factors influencing fruit cracking: an environmental and agronomic perspective

Fruit cracking, a widespread physiological disorder affecting various fruit crops and vegetables, has profound implications for fruit quality and marketability. This mini review delves into the multifaceted factors contributing to fruit cracking and emphasizes the pivotal roles of environmental and agronomic factors in its occurrence. Environmental variables such as temperature, relative humidity, and light exposure are explored as determinants factors influencing fruit cracking susceptibility. Furthermore, the significance of mineral nutrition and plant growth regulators in mitigating fruit cracking risk is elucidated, being calcium deficiency identified as a prominent variable in various fruit species. In recent years, precision farming and monitoring systems have emerged as valuable tools for managing environmental factors and optimizing fruit production. By meticulously tracking parameters such as temperature, humidity, soil moisture, and fruit skin temperature, growers can make informed decisions to prevent or alleviate fruit cracking. In conclusion, effective prevention of fruit cracking necessitates a comprehensive approach that encompasses both environmental and agronomic factors

Comparison of a clinic-based ELISA test kit with the immunofluorescence antibody test for assaying Leishmania infantum antibodies in dogs

This study compares a rapid Immunospecific Kalazar Canine Rapid Spot IF with the gold standard test (indirect fluorescent antibody test (IFAT)) for detection of Leishmania infantum specific IgG serum antibodies in naturally exposed dogs. Serum samples were obtained from 89 healthy dogs and dogs affected by canine leishmaniosis (CanL). IgG-IFAT titers ≥80 were considered positive. Anti-L. infantum IgG antibodies were found in 54 samples with titers ranging from 1 : 80 to 1 : 5120. The performance of the rapid Immunospecific Kalazar was evaluated using a ROC curve. The area under the ROC curve of 0.957 was significantly different from 0.5 ( < 0.0001), and therefore it can be concluded that the rapid Immunospecific Kalazar has the ability to distinguish canine sera with and without L. infantum IgG. The best performance of the test was at a cutoff >0 (sensitivity 92.6%, specificity 97%). The test can be used for disease screening if the cutoff is >0 (highest sensitivity, 92.6%) and is recommended as confirmatory test for the presence of L. infantum IgG antibodies if the cutoff is set >2 (highest specificity, 100%)

Frecuencia del grupo sanguíneo A, B y AB y riesgo de transfusión en gatos del Norte de Italia

Objetivos Datos epidemiol\uf3gicos sobre el tipo de sangre felina son importantes para prevenir reacciones de transfu- si\uf3n y para identificar los donantes de sangre ideales en programas de donantes de sangre felina. Tres eran los objetivos de este estudio: 1. Determinarlafrecuenciadelosgrupossangu\uedneos en una poblaci\uf3n de gatos con tipaje sangu\uedneo en la Universidad de Mil\ue1n 2. Analizar las caracter\uedsticas de la poblaci\uf3n y pro- bar la asociaci\uf3n entre el tipo de sangre A, B y AB y una serie de variables (sexo, raza y grupo filoge- n\ue9tico); 3. Calcular el riesgo de reacci\uf3n a la transfusi\uf3n menor y mayor despu\ue9s de la administraci\uf3n de sangre incompatible despu\ue9s de una transfusi\uf3n al azar en esta poblaci\uf3n. 4. Calcular el riesgo de una reacci\uf3n a latransfusi\uf3n mayor o menor despu\ue9s de una administraci\uf3n de sangre incompatible de forma aleatoria en esta poblaci\uf3n Material y m\ue9todos Un estudio retrospectivo se realiz\uf3 utilizando datos (recogidos entre 2010 y 2014) de la base de datos de la unidad de transfusi\uf3n veterinaria (REV) de la Universidad de Mil\ue1n, Italia. Los datos recogidos inclu\uedan: tipo de sangre, edad, sexo, raza, raz\uf3n y el m\ue9todo de determinaci\uf3n del grupo sangu\uedneo. Los resultados se analizaron por an\ue1lisis de frecuencia absoluta y relativa. Las frecuencias de los diferentes grupos sangu\uedneos y datos categ\uf3ricos (sexo, raza y grupo filogen\ue9tico) se compararon mediante tablas de contingencia y la prueba exacta de Fisher o prueba de chi cuadrado, seg\ufan el caso. Se seleccionaron Conclusiones 88 gatos como potenciales donantes de sangre, el grupo sangu\uedneo y otros factores (precruce, preoperatorio y receptor) fueron considerados aptos para el objetivo de los c\ue1lculos. La posibilidad de una reacci\uf3n mayor de transfusi\uf3n en gato con sangre del grupo B que recibe sangre del tipo A o AB, se calcul\uf3 de la siguiente manera: frecuencia del donante tipo A + frecuencia d\uf3nate tipo AB multiplicada ambas por frecuencia de receptor B. La posibilidad de una reacci\uf3n de transfusi\uf3n menor (la reacci\uf3n menor reduce la vida media de los eritrocitos transfundidos en sangre de gatos del grupo sangu\uedneo A que recibe sangre del grupo sangu\uedneo B o AB y gatos del grupo sangu\uedneo AB que reciben sangre de grupo B) fue calculada usando la f\uf3rmula: frecuencia del gato donante tipo B donante + frecuencia del donante tipo AB multiplicado por la frecuencia del receptor tipo A + frecuencia donante tipo B multiplicado por frecuencia receptor tipo AB (Juvet F et al 2011). Resultados 282 fueron incluidos en el estudio, el rango de edad fue desde 2 meses a 19 a\uf1os, 140 (50%) macho y 138 (49%) hembras, 151 (53%) gato com\ufan europeo de pelo corto, 59 (21%) Ragdoll, 29 (10%) Maine Coons. Los gatos se dividieron en tres grupos de acuerdo con su origen 55% de los gatos eran de la cuenca medite- rr\ue1nea, 44% del oeste de Europa, 1% de Asia y no se tipificaron gatos de \uc1frica oriental. La prevalencia de los grupos sangu\uedneos fue 91% (n= 257) tipo A, 4% (n=11) tipo B y 5% (n=14) tipo AB. Las razones para realizar el grupo sangu\uedneo fueron: precruce para pre- venir Isoeritrolisis neonatal en 92 gatos (33%), scree- ning preoperatorio en 55 gatos (19%), examen pre- transfusi\uf3n en 17 gatos (6%) y screening de potenciales donantes previo a la donaci\uf3n en 88 gatos (31%). No hubo asociaci\uf3n significativa entre el grupo sangu\uedneo y la variable analizada, excepto por una asociaci\uf3n entre gato Ragdoll y grupo sangu\uedneo A (83%) y AB (14%) (P=0,0335, OR=0,3 y P=0,0026, OR=5,6 respecti- vamente). Todos los gatos Maine Coon testado fueron grupo sangu\uedneo A. En una transfusi\uf3n de sangre sin testar la frecuencia estimada de una reacci\uf3n mayor y menor fue 3.8% y 9.3% respectivamente. Discusi\uf3n De acuerdo con los hallazgos de otros estudios europeos (Jensen AL et al, 1994; Knottembelt CM et al 1999; Ruiz de Gopegui R et al 2004; Marques C et al 2011; Juvet F et al, 2011), el grupo sangu\uedneo predominante en los gatos en este estudio fue de tipo A. La prevalencia del grupos sangu\uedneo var\ueda entre las diferentes razas felinas. El tipo de sangre AB es significativamente m\ue1s frecuente en los gatos de Ragdoll que en otras razas feli- nas y este hallazgo podr\ueda ser \ufatil cuando se consideran candidatos para la donaci\uf3n de plasma. Conclusiones Dado que el riesgo de una reacci\uf3n transfusional en los gatos que recibieron una transfusi\uf3n de sangre sin la comprobaci\uf3n de la compatibilidad sangu\uednea que resulto ser del 9% el grupo sangu\uedneo y las pruebas cruzadas de los donantes de sangre y el receptor son esenciales antes de la transfusi\uf3n felina para prevenir la potencial reacci\uf3n transfusional inmunol\uf3gica o la r\ue1pida destrucci\uf3n de los eritrocitos transfundidos. Bibliograf\ueda \u2022 jensen AL, Olesen AB, Arnbjerg J. Distribution of feli- ne blood types obtained in the Copenhagen area of Den- mark. Acta Vet Scand. 1994;35:121-4. \u2022 Knottenbelt CM, Addie DD, Day MJ, Mackin AJ. Deter- mination of the prevalence of feline blood types in the UK. J Small Anim Pract. 1999;40:115-8. \u2022 Ruiz de Gopegui R, Velasquez M, Espada Y. Survey of fe- line blood types in the Barcelona area of Spain. Vet Rec. 2004;154:794-5. \u2022 Marques C, Ferreira M, Gomes JF. Frequency of blood type A, B and AB in 515 domestic shorthair cats from the Lisbon area. Vet Clin Pathol. 2011;40:185-7. \u2022 Juvet F, Brennan S, Mooney CT. Assessment of feline blood for transfusion purposes in the Dublin area of Ireland. Vet Rec. 2011;168:352-4