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

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

Use of Autologous Leucocyte- and Platelet-Rich Plasma (L-PRP) in the Treatment of Aural Hematoma in Dogs

Leukocyte- and platelet-rich plasma (L-PRP) can accelerate the healing process by providing increased concentrations of platelet-derived growth factors. The objective of this study was to evaluate the clinical effect of L-PRP in the treatment of canine aural hematomas associated with otitis externa. Twenty mL of citrated whole blood was collected from each of the 17 dogs included and autologous L-PRP was produced. The aural hematoma was drained and 0.5–1 mL of L-PRP was injected. The dogs were examined weekly until 7 days after complete clinical healing. A final clinical follow-up was performed 6 weeks after the first treatment with L-PRP. If there was recurrence of the aural hematoma at the first follow-up, the treatment was repeated. In total, 2/17 cases were lost after the first follow-up. In 5/17 dogs, a short-term recurrence occurred. In 12/15 cases, complete clinical resolution was achieved with a single L-PRP application (Group A1) and in 3/15 with two treatments (Group A2). The mean time to complete clinical resolution was 16 ± 8.7 days (A1) and 23.3 ± 4 days (A2), respectively. No side effects were reported. The in situ administration of autologous L-PRP resulted in a complete resolution of the aural hematoma in all dogs that completed the clinical trial

Efficacy of a Semi Automated Commercial Closed System for Autologous Leukocyte- and Platelet-Rich Plasma (l-prp) Production in Dogs: A Preliminary Study

Background: To characterize the cellular composition (platelets, erythrocytes, and leukocytes) and determine platelet-derived growth factor isoform BB (PDGF-BB) concentration in canine leukocyte- and platelet rich plasma (L-PRP) produced using a commercial semi-automated closed system. Methods: Twenty milliliters of citrated whole blood were obtained from 30 healthy un-sedated canine blood donors and processed using a semi-automated completely closed commercial system (CPUNT 20, Eltek group, Casale Monferrato, Alessandria, Italy) according to the manufacturer’s instructions. Erythrocyte, leukocyte, and platelet counts were determined in both whole blood (WB) and resultant L-PRP. The PDGF-BB concentration was evaluated after bovine thrombin activation of 10 L-PRP samples. Results: This commercial system produced on average 2.3 ± 0.7 mL of L-PRP containing a high concentration of platelets (767,633 ± 291,001 μL, p < 0.001), with a 4.4 fold increase in platelet count, lower concentration of erythrocytes (528,600 ± 222,773 μL, p < 0.001) and similar concentration of leukocytes (8422 ± 6346 μL, p = 0.9918) compared with WB. L-PRP had an average of 3442 ± 2061 pg/mL of PDGF-BB after thrombin activation. Neutrophils, lymphocytes and monocytes average percent content in L-PRP was 14.8 ± 13.2, 71.7 ± 18.5 and 10.7 ± 6.4, respectively. Conclusion: Sterile canine L-PRP prepared using this semi-automated closed system is easy to obtain, produces a significant increase in platelet count compared to WB and contains a detectable concentration of PDGF-BB after activation. Additional in vitro and in vivo studies are needed to assess inflammatory markers concentration and the therapeutic efficacy of this L-PRP in dogs

Evaluation of Association between Blood Phenotypes A, B and AB and Feline Coronavirus Infection in Cats

Cats are susceptible to feline coronavirus (FCoV), a highly contagious virus with fecal–oral transmission. In people, susceptibility to coronavirus infection, such as SARS-CoV infection, has been associated with the ABO blood group, with individuals with blood group O having significantly lower risk of SARS-CoV infection. This study evaluated a possible association between feline blood group phenotypes A, B and AB and serostatus for antibodies against FCoV. We also investigated risk or protective factors associated with seropositivity for FCoV in the investigated population. Feline populations were surveyed for AB group system blood types and for presence of antibodies against FCoV. Blood phenotype, origin, breed, gender, reproductive status and age of cats were evaluated as protective or risk factors for coronavirus infection. No blood type was associated with FCoV seropositivity, for which being a colony stray cat (p = 0.0002, OR = 0.2, 95% CI: 0.14–0.54) or a domestic shorthair cat (p = 0.0075, OR = 0.2, 95% CI = 0.09–0.69) were protective factors. Based on results of this study, feline blood phenotypes A, B or AB do not seem to predispose cats to seropositivity for FCoV. Future studies on other feline blood types and other infections could clarify whether feline blood types could play a role in predisposing to, or protecting against, feline infections

Serum Protein Gel Agarose Electrophoresis in Captive Tigers

Given the endangered status of tigers (Panthera tigris), the health of each individual is important and any data on blood chemistry values can provide valuable information alongside the assessment of physical condition. The nature of tigers in the wild makes it is extremely difficult to obtain biological samples from free-living subjects, therefore the values obtained from captive tigers provide very useful data. Serum protein electrophoresis is a useful tool in the diagnosis and monitoring of a number of diseases. In this study, we evaluated agarose gel serum protein electrophoresis on samples from 11 healthy captive tigers. Serum electrophoresis on all 11 tiger samples successfully separated proteins into albumin, α1, α2, β1, β2 and γ globulin fractions as in other mammals. Electrophoretic patterns were comparable in all tigers. Mean± standard deviation or median and range values obtained for each protein fraction in healthy tigers were, respectively: 3.6 ± 0.2, 0.21 (0.2–0.23), 1.2 ± 0.2, 10.7 ± 0.2, 0.4 (0.3–0.6), 1.2 (1–1.8) gr/dL. The results of this preliminary study provide the first data on serum electrophoretic patterns in tigers and may be a useful diagnostic tool in the health assessment of this endangered species

Neuroprotective Nanoparticles Targeting the Retina: A Polymeric Platform for Ocular Drug Delivery Applications

Neuroprotective drug delivery to the posterior segment of the eye represents a major challenge to counteract vision loss. This work focuses on the development of a polymer-based nanocarrier, specifically designed for targeting the posterior eye. Polyacrylamide nanoparticles (ANPs) were synthesised and characterised, and their high binding efficiency was exploited to gain both ocular targeting and neuroprotective capabilities, through conjugation with peanut agglutinin (ANP:PNA) and neurotrophin nerve growth factor (ANP:PNA:NGF). The neuroprotective activity of ANP:PNA:NGF was assessed in an oxidative stress-induced retinal degeneration model using the teleost zebrafish. Upon nanoformulation, NGF improved the visual function of zebrafish larvae after the intravitreal injection of hydrogen peroxide, accompanied by a reduction in the number of apoptotic cells in the retina. Additionally, ANP:PNA:NGF counteracted the impairment of visual behaviour in zebrafish larvae exposed to cigarette smoke extract (CSE). Collectively, these data suggest that our polymeric drug delivery system represents a promising strategy for implementing targeted treatment against retinal degeneration