Serum Biochemistry Parameters of the Endangered Miranda's Donkey Breed: Reference Intervals and the Influence of Gender and Age.

Miranda's donkey is an endangered, autochthone breed from Northern Portugal. Understanding the physiological and pathological conditions of Miranda's donkey is crucial for the conservation of the breed. Our study aimed to establish reference intervals (RIs) for blood biochemistry parameters and to evaluate the influence of gender and age on these parameters. Blood samples from 75 clinically healthy animals were analyzed for 21 blood biochemistry parameters using Respons® 920 and Start® 4-Diagnostica-Stago. RIs were calculated according to the ASVCP guidelines, utilizing Reference Advisor software V. 2.1 and the statistical program SPSS version 29 to analyze the effects of gender and age. Significant gender-related differences (p < 0.05) were observed in cholesterol, chlorine, creatinine (CREA), glucose (GLU), sodium, and triglycerides (with higher values in females) and in aspartate aminotransferase, creatine phosphokinase (CK), gamma-glutamyl transferase, potassium, magnesium (Mg), and urea (with higher values in males). Age-related differences (p < 0.05) were noted for alkaline phosphatase, CK, fibrinogen, GLU, and phosphorus (higher in young animals) and for globulin, Mg, and total protein (higher in adults). The RIs described here are invaluable for assessing and monitoring the health status of individual animals and herds. Our study highlights the importance of considering gender and age in the interpretation of biochemical parameters, providing critical insights for the conservation and management of this endangered breed.

A Review on Blood Reference Values as a Valuable Marker of Wildlife Welfare in Erinaceus europaeus.

The western-European hedgehog (Erinaceus europaeus), in expanding its range towards human habitats, faces exposure to contaminants and biological agents, potentially leading to diseases associated with hematological and biochemical changes. As bioindicators of environmental pollution and carriers of zoonotic agents, hedgehogs play a crucial role in One Health studies, emphasizing the need for a comprehensive understanding of their clinical-pathological aspects. Exploring the blood reference values in healthy animals of this species is crucial for understanding and improving their well-being, and identifying possible diseases/pathogens that may affect its conservation and/or impact human health. This review is focused on analyzing the data available in the literature for Erinaceus europaeus blood reference intervals. A comprehensive literature review of the studies published in Europe is performed, highlighting their specificities, and emphasizing the need for continuous research in this field. Our final goal is to provide a crucial tool for assessing the health status of the species, and underscoring the significance of research in this specific domain.

Establishment of Reference Intervals of Hematological Parameters and Evaluation of Sex and Age Effect in the Miranda Donkey.

The Miranda donkey is an autochthonous Portuguese breed that is considered endangered. Several studies have been carried out on this breed, but to the authors' best knowledge, no studies have been conducted on their clinical pathology. The aims of this study were to determine the hematological reference intervals (RIs) in healthy Miranda donkeys and to estimate the influence of age and sex. Blood samples from 75 clinically healthy animals were analyzed for 22 hematological parameters on the IDEXX ProCyte Dx, an automated hematology analyzer previously validated for the species. The RIs were estimated following the ASVCP guidelines with the Reference Value Advisor software. Regarding sex, no significant differences were found between groups. Regarding age, significant statistical differences (p < 0.05) were observed for red blood cells, red cell distribution width, white blood cells, lymphocytes, monocytes, platelets, plateletcrit (higher mean in young animals), mean corpuscular volume, mean corpuscular haemoglobin, neutrophils and eosinophils (higher mean in adults). The RIs described here can be used to evaluate and monitor the health status of animals and herds, as well as to guide diagnoses or select fit and healthy animals for reproduction, contributing to the preservation of the breed.

Hematology, Biochemistry, and Protein Electrophoresis Reference Intervals of Western European Hedgehog (Erinaceus europaeus) from a Rehabilitation Center in Northern Portugal.

The Western European hedgehog (Erinaceus europaeus) can work as a bioindicator of environmental pollution and be a host for multiple zoonotic agents, making it relevant in terms of One Health studies. It is essential to deepen the knowledge on this species and calculate reference intervals (RIs) for the usual hematological and biochemical parameters. For this retrospective study (2017-2022), the archives of the Clinical Pathology Laboratory (LPC) of University of Trás-os-Montes and Alto Douro (UTAD) Veterinary Teaching Hospital were analyzed. Data of hematology, clinical biochemistry, and protein electrophoresis from 37 healthy hedgehogs of the Wild Animal Rehabilitation Center at UTAD, Northern Portugal, were included. It was possible to calculate RIs for almost all of the variables in the study, using Reference Value Advisor V2.1. Moreover, sex and age effects were investigated: alkaline phosphatase (p = 0.012, higher in males); total proteins (p = 0.034, higher in adults); mean cell volume (p = 0.007) and mean corpuscular hemoglobin (p = 0.010) (both higher in juveniles); and red blood cell distribution width (p = 0.021, higher in adults). Our study allowed for the first time to define RIs for a population of hedgehogs in Portugal, having a potentially relevant impact on species conservation and in the human-animal health interface.

Prothrombin time and activated partial thromboplastin time reference intervals in dogs, by gender and age group using the Start®4 (Stago) / Intervalos de referência para o tempo de protrombina e tempo de tromboplastina parcial ativada no cão de acordo com a idade e o gênero utilizando o Start ® 4 (Stago)

Prothrombin time (PT) and the activated partial thromboplastin time (aPTT) are useful tools for the diagnosis and monitoring of coagulation disorders in Veterinary Medicine. Our objectives were: to establish reference intervals (RI) for PT and a PTT for the dog using the Start®4 (Stago), to compare the obtained RI with literature; to evaluate the effects of gender and age on the coagulation profile. Plasma samples of 122 healthy dogs (57 males; 65 females) aged between 4 months and 18 years, divided into three age groups (0-2 years old; 3-10 years old; > 10 years old) and grouped in to males and females were analysed. The RI were estimated following the ASVCP guidelines with the Reference Value Advisor software. The RI were: PT 6.7'' to 10.8''; aPTT 9.0'' to 14.8''. PT was significantly higher in females than in males. Dogs aged 10 years or older have significantly higher mean aPTT times than younger dogs. RI comparison showed a considerable percentage of cases outside the reference RI of the literature (PT - 79.3%; aPTT - 77.1%), demonstrating the need of each laboratory to calculate its own RI. The RI established in this study are applicable for the coagulation profile assessment in dogs.

O tempo de protrombina (TP) e o tempo de tromboplastina parcial ativada (TTPa) são ferramentas úteis para o diagnóstico e monitorização das alterações da coagulação em Medicina Veterinária. Os objetivos deste estudo foram: estabelecer intervalos de referência (IR) para TP e TTPa para o cão utilizando o Start®4 (Stago), de modo a comparar os IR obtidos com a literatura; avaliar os efeitos do sexo e da idade no perfil da coagulação. Foram usadas amostras de plasma de 122 cães saudáveis (57 machos; 65 fêmeas) com idades entre quatro meses e 18 anos, divididos em três grupos (0-2 anos; 3-10 anos; > 10 anos) e agrupados em machos e fêmeas. Os IR foram calculados seguindo as diretrizes da ASVCP com o software Reference Value Advisor. Os IR obtidos foram: PT 6,7 '' a 10,8 ''; TTPa 9,0 '' a 14,8 ''. O TP foi significativamente maior nas fêmeas do que nos machos. Os cães com 10 anos ou mais apresentaram tempos médios de TTPa significativamente maiores do que cães mais jovens. A comparação de IR mostrou uma percentagem considerável de casos fora do IR de referência da literatura (TP - 79,3%; TTPa - 77,1%), confirmando a necessidade de cada laboratório calcular seu próprio IR. Os IR estabelecidos neste estudo são aplicáveis na avaliação do perfil hemostático em cães.

Acute phase proteins and biomarkers of oxidative status in feline spontaneous malignant mammary tumours.

Acute phase proteins (APP) and biomarkers of oxidative status change in human and canine mammary tumours, however, they have not been studied in feline mammary tumours. The aims of this study were to investigate the APP and antioxidant responses in feline malignant mammary tumours, to evaluate their relation with tumour features, and to assess their prognostic value. Serum amyloid A (SAA), haptoglobin (Hp), albumin, butyrylcholinesterase (BChE), insulin-like growth factor1 (IGF1), paraoxonase1 (PON1), total serum thiols (Thiol), glutathione peroxidase (GPox) and total antioxidant capacity determined by different assays, including trolox equivalent antioxidant capacity assessed by two different methodologies (TEAC1/2), ferric reducing ability of plasma (FRAP), and cupric reducing antioxidant capacity (CUPRAC), were determined in serum of 50 queens with spontaneous mammary carcinomas and of 12 healthy female cats. At diagnosis, diseased queens presented significantly higher SAA and Hp, and lower albumin, BChE, GPox, TEAC1, TEAC2 and CUPRAC than controls. Different tumour features influenced concentrations of APP and antioxidants. Increases in serum Hp, and decreases in albumin, Thiol and FRAP were significantly associated with neoplastic vascular emboli, metastasis in regional lymph nodes and/or in distant organs. Distant metastasis development during the course of the disease was associated with increases in SAA and TEAC1. At diagnosis, decreased albumin was associated with a longer survival, and BChE <1.15 µmoL/mL.minute was associated with a shorter survival time on multivariate analysis. Feline malignant mammary tumours are associated with an APP response and oxidative stress, and different tumour features influence the inflammatory response and the oxidative damage. Furthermore, some of these analytes proved to have prognostic value.

Acute phase proteins and antioxidant responses in queens with pyometra.

Acute phase proteins (APP) and biomarkers of oxidative status have proved to be clinically useful biomarkers of pyometra in different species. Despite pyometra is considered one of the most important feline reproductive diseases, information about the APP response and the oxidative status in queens with pyometra is lacking. This study aimed to evaluate the APP and the antioxidant responses at diagnosis and in the post-operative period in feline pyometra. Serum concentrations of serum amyloid A (SAA), haptoglobin (Hp), albumin, total serum thiols (Thiol) and total antioxidant capacity determined by different assays, including trolox equivalent antioxidant capacity (TEAC) assessed by two different methodologies (TEAC1 and TEAC2), ferric reducing ability of plasma (FRAP), and cupric reducing antioxidant capacity (CUPRAC), were determined in 23 queens with pyometra at diagnosis and in 13 healthy control queens submitted to elective ovariohysterectomy. The APP and antioxidants were also evaluated in 11 queens of the pyometra group at days two and 10 after surgery. At diagnosis, queens with pyometra had serum concentrations of SAA, Hp, and FRAP significantly higher (P < 0.001, P < 0.001 and P < 0.05, respectively), and of albumin, Thiol, CUPRAC and TEAC2 significantly lower (P < 0.001, P < 0.001, P < 0.001 and P < 0.01, respectively) than controls. Moreover, concentrations of APP and antioxidants were significantly different (with a tendency to return to physiologic levels) at day 10 after surgery than before surgery. Significant associations were found between APP and antioxidants. According to these results, an APP response and the development of oxidative stress were detected in queens with pyometra. In addition, APP and antioxidants tended to return to physiologic values after surgery in the queens that recovered from the disease. Therefore, our results suggest that APP and selected antioxidants, such as Thiol and CUPRAC, could be potentially useful biomarkers in diagnosis and assessment of the post-operative period in feline pyometra.

Acute phase proteins response in cats naturally infected by hemotropic mycoplasmas.

Information about the acute phase proteins (APP) response in cats naturally infected with hemoplasmas and in cats co-infected with different species of hemoplasmas is lacking. This study evaluated serum amyloid A (SAA), haptoglobin (Hp) and albumin in 48 cats naturally infected with hemoplasmas, including 25 with Candidatus Mycoplasma haemominutum and 23 co-infected with different hemoplasmas agents; and in 10 healthy control cats. Infected cats had significantly higher Hp and lower albumin than controls. Symptomatic cats had significantly higher SAA and Hp, and lower albumin than asymptomatic animals, and also than controls. Asymptomatic cats had significantly higher Hp than controls. Concentrations of APP were not significantly different between single infected and co-infected cats. According with these results, hemoplasmosis should be considered when alterations in APP are detected in diseased cats with compatible clinical signs. Furthermore, a subclinical infection should be considered in apparently healthy cats from endemic areas with increased Hp.

Acute phase proteins response in cats naturally infected with Hepatozoon felis and Babesia vogeli.

BACKGROUND: The measurement of acute phase proteins (APP) is being increasingly used in human and veterinary medicine in diagnosis, prognosis, treatment monitoring, and in general health screening. However, information about the APP response in cats infected with agents of vector-borne diseases is lacking. OBJECTIVES: The purpose of the study was to investigate the concentrations of serum amyloid A (SAA), haptoglobin (Hp), and paraoxonase-1 (PON1) in cats naturally infected with Hepatozoon felis and Babesia vogeli. METHODS: Serum concentrations of SAA, Hp, and PON1 were determined in 19 cats naturally infected with H felis and in 11 cats naturally infected with B vogeli, and compared to concentrations in 10 healthy control cats. RESULTS: Serum Hp concentrations were significantly increased, and PON1 concentrations significantly decreased in symptomatic and asymptomatic cats infected with H felis and B vogeli when compared with healthy noninfected cats. In the H felis-infected population, concentrations of SAA and Hp were significantly increased in symptomatic cats when compared with asymptomatic animals. CONCLUSIONS: This study demonstrated differences in APP concentrations in cats infected with H felis and B vogeli. Therefore, Hp and PON1 concentrations could be helpful in discriminating healthy cats from cats with asymptomatic or symptomatic infection by these agents.

Serum acute phase proteins in Dirofilaria immitis and Wolbachia seropositive cats.

Objectives The aim of this study was to characterise the response of acute phase proteins (APPs) in cats seropositive for Dirofilaria immitis and to its endosymbiont bacterium Wolbachia. Methods The APPs serum amyloid A (SAA), haptoglobin (Hp) and ceruloplasmin (Cp) were measured in 25 seropositive cats and in 16 healthy seronegative cats. Results SAA and Cp concentrations were significantly higher in animals with D immitis seropositivity that exhibited clinical signs related to the disease, and Hp was elevated in all D immitis-seropositive animals. There was no significant correlation between APPs and D immitis or Wolbachia species antibody titres. Conclusions and relevance An association between feline seropositivity to D immitis and APP response was demonstrated. Increases in serum SAA and Cp concentrations were related to D immitis-associated clinical signs, whereas Hp increased in all seropositive animals.

Urine protein-to-creatinine concentration ratio in samples collected by means of cystocentesis versus manual compression in cats.

Objective-To compare urine protein-to-creatinine concentration (UPC) ratios in samples collected by means of cystocentesis versus manual compression in cats. Design-Evaluation study. Animals-43 client-owned cats requiring urinalysis. Procedures-In all cats, 5 mL of urine from the midstream phase of micturition was collected by means of manual compression and, subsequently, an additional 5 mL of urine was obtained by means of ultrasound-guided cystocentesis. A complete urinalysis was performed on all samples, and UPC ratios were determined. Results-Cats were classified on the basis of the International Renal Interest Society substaging system as being free from proteinuria (UPC ratio, < 0.2; n = 19) or as having borderline proteinuria (UPC ratio, 0.2 to 0.4; 7) or proteinuria (UPC ratio, > 0.4; 17). None of the cats had postrenal proteinuria. A significant linear correlation was identified between UPC ratios in urine samples obtained by means of manual compression and ratios in samples obtained by means of cystocentesis. For all cats, UPC ratios for samples obtained by the 2 collection methods resulted in classification in the same IRIS substage. Conclusions and Clinical Relevance-Results suggested that collection of a urine sample from the midstream phase of micturition by manual compression would be a reliable alternative to cystocentesis for the determination of UPC ratio in cats, provided that postrenal proteinuria was excluded by means of urine sediment analysis. Once postrenal proteinuria was ruled out, the method used to collect urine samples did not appear to influence the quantification of urine protein concentration.

Feline vector-borne pathogens in the north and centre of Portugal.

BACKGROUND: In recent years, several clinical cases and epidemiological studies of feline vector-borne diseases (FVBD) have been reported worldwide. Nonetheless, information on FVBD agents and their prevalence in Portugal is scarce. METHODS: Three-hundred and twenty domestic cats presented to 30 veterinary medical centres in the north and centre regions of Portugal were randomly sampled. Blood was assayed by real-time polymerase chain reaction (PCR) for genera Anaplasma/Ehrlichia, genus Babesia, Hepatozoon canis, Hepatozoon felis, Leishmania infantum and the genus Rickettsia. Babesia-positive samples were further tested for Babesia canis and Babesia vogeli. RESULTS: Eighty (25.0%) out of the 320 cats were positive to at least one vector-borne agent, including seven (2.2%) cats co-infected with two agents. Two cats (0.6%) were infected with Anaplasma/Ehrlichia spp., four (1.3%) with B. canis, 26 (8.1%) with B. vogeli, 50 (15.6%) with H. felis, one (0.3%) with L. infantum and four (1.3%) with Rickettsia spp. No cat tested positive for H. canis. One cat (0.3%) was co-infected with B. canis and B. vogeli, three (0.9%) with B. vogeli and H. felis, one (0.3%) with H. felis and L. infantum, and two (0.6%) with H. felis and Rickettsia spp. CONCLUSIONS: A considerable prevalence of infection with vector-borne pathogens among the domestic feline population of the north and centre of Portugal has been revealed by the present study. Additionally, this is the first detection of B. vogeli in cats from Europe and of H. felis in cats from Portugal.

Whole blood transfusion in small animals: indications and effects.

Transfusion therapy is a major resource that can improve the patient's capability to overcome the underlying disease. However, the effects of whole blood infusion, and how they affect the patient's outcome, are not yet clear. For this study, a protocol was developed in order to monitor a group of 15 animals (9 dogs, 6 cats) that received a total of 19 transfusions; 3 animals received more than one transfusion each. The most common indications for blood transfusion included acute blood loss (47%), coagulopathy (33%) and other anaemias (20%). The mean pre-transfusion packed cell volume (PCV) of animals with acute blood loss (18%) was higher than in the group of coagulopathy (15%) or other anaemias (15%). The survival rates at 6 days after transfusion were greater in the coagulopathy (80.0%) and other anaemias (66.7%) than in the group of acute blood loss (42.9%). After transfusion, pulse rate ( p <0.01) and platelet count ( p <0.05) decreased significantly, and there was a significant increase in body temperature of the animals that suffered from hypothermia before the transfusion ( p <0.05). Overall survival was predictable based upon posttransfusion body temperature, observed PCV change, the difference between the obtained and the calculated PCV, and administered transfusion volume ( p <0.05).

Whole blood transfusion in small animals: indications and effects

Transfusion therapy is a major resource that can improve the patient's capability to overcome the underlying disease. However, the effects of whole blood infusion, and how they affect the patient's outcome, are not yet clear. For this study, a protocol was developed in order to monitor a group of 15 animals (9 dogs, 6 cats) that received a total of 19 transfusions; 3 animals received more than one transfusion each. The most common indications for blood transfusion included acute blood loss (47 percent), coagulopathy (33 percent) and other anaemias (20 percent). The mean pre-transfusion packed cell volume (PCV) of animals with acute blood loss (18 percent) was higher than in the group of coagulopathy (15 percent) or other anaemias (15 percent). The survival rates at 6 days after transfusion were greater in the coagulopathy (80.0 percent) and other anaemias (66.7 percent) than in the group of acute blood loss (42.9 percent). After transfusion, pulse rate ( p <0.01) and platelet count ( p <0.05) decreased significantly, and there was a significant increase in body temperature of the animals that suffered from hypothermia before the transfusion ( p <0.05). Overall survival was predictable based upon posttransfusion body temperature, observed PCV change, the difference between the obtained and the calculated PCV, and administered transfusion volume ( p <0.05).

A terapia transfusional é um importante recurso que pode auxiliar na recuperação do doente face à patologia primária. No entanto, não estão totalmente esclarecidos os efeitos de uma transfusão sanguínea, e que consequências podem ter no desfecho clínico. Para este estudo, foi desenvolvido um protocolo de forma a monitorizar 15 animais (9 cães e 6 gatos) que receberam um total de 19 transfusões; 3 animais receberam mais do que uma unidade de sangue. As indicações mais comuns para realização da transfusão foram hemorragia aguda (47 por cento), coagulopatia (33 por cento) e outras anemias (20 por cento). O hematócrito pré-transfusional médio dos animais com hemorragia aguda (18 por cento) foi superior ao dos animais com coagulopatia (15 por cento) ou outras anemias (15 por cento). A taxa de sobrevivência ao sexto dia pós-transfusão foi maior no grupo da coagulopatia (80,0 por cento) e de outras anemias (66,7 por cento), do que no grupo da hemorragia aguda (42,9 por cento). Após a transfusão, a frequência de pulso ( p <0,01) e a contagem de plaquetas ( p <0,05) diminuíram significativamente, e houve um aumento significativo da temperatura corporal nos animais com hipotermia antes da transfusão ( p <0,05). A sobrevivência foi predizível com base na temperatura pós-transfusão, a variação do Ht, a diferença entre o Ht real e o Ht esperado, e o volume de transfusão aplicado ( p <0,05).

Feline neonatal isoerythrolysis and the importance of feline blood types.

Although feline neonatal isoerythrolysis is rare, associated mortality rate is high. It results from mating of type B blood queens with type A or AB blood toms. A comprehensive review on feline blood types and feline neonatal isoerythrolysis physiopathology, clinical features, diagnosis, treatment, and prevention is covered.

Frequencies of feline blood types in northern Portugal.

The frequencies of feline blood types in northern Portugal were studied by surveying 185 pedigreed and nonpedigreed cats. Blood typing was performed by the traditional tube method. As a single group, the majority of cats were type A (90.3%), 3.8% were type B, and 5.9% were type AB. Among pedigreed cats, 19 were Siamese and 7 were Persian; all but 1 were type A. Among nonpedigreed cats, 89.3% were type A, 4.4% were type B, and 6.3% were type AB.