Elizabeth Moreira dos Santos Schmidt

Hematology of the Red-capped parrot (Pionopsitta pileata) and Vinaceous Amazon parrot (Amazona vinacea) in captivity.

Abstract Preliminary reference intervals for hematologic and total plasma protein profiles were determined for nine adult Red-capped parrots (Pionopsitta pileata) (six males and three females) and six Vinaceous Amazon parrots (Amazona vinacea) (two adult males, two adult females, one juvenile, and one nonsexed) from the Curitiba Zoo, Paraná, Brazil. For both Red-capped parrots and Vinaceous Amazon parrots, adult males had higher red blood cell counts than adult females. Regarding white blood cell distribution, differences due to gender were also found for both species of parrots.

The final hurdles for acute phase protein analysis in small animal practice

Studies on the acute phase proteins (APP) in small animal medicine have proliferated recently with particular emphasis on their value as biomarkers of infectious, inflammatory and neoplastic diseases. There have been several reviews on the extensive knowledge that has thus accumulated (Eckersall & Conner 1988, Ceron et al. 2005, Eckersall & Bell 2010, Cray et al. 2009). However, in many parts of the world the majority of veterinary practitioners do not have tests for APPs available on a regular basis. A number of pioneering laboratories have established methods that can be used for routine analysis for small animal practice (Jensen & Kjelgaard-Hansen 2006, Caldin et al. 2009, Martinez-Subiela et al. 2003, Shida et al. 2011). For less specialised laboratories there are significant problems to be overcome prior to wide spread availability of APP assays. Notable are the lack of a universally accepted calibration standard and the availability of a quality assurance (QA) scheme to ensure commonality between reported results. Here we review selected recent publications on APP in companion animals and thereafter discuss how to overcome such hurdles to the full use of APP assays in these species. Only a few studies have evaluated temporal changes of APP in companion animals. In one such study, dogs with severe clinical signs of parvovirus enteritis that did not survive had higher C-reactive protein (CRP) concentrations at admission, and at 12 and 24 hours after admission when compared with the dogs that survived. Higher CRP was also associated with longer hospitalisation time and the sensitivity and specificity of CRP to differentiate between survivors and non-survivors at 24 hours after admission was 86·7 and 78·7%, respectively (McClure et al. 2013). In a complementary investigation, dogs that did not survive the infection and had leukopenia and also severe clinical signs of parvovirus enteritis had a 72-fold increase in CRP and a 3·1-fold increase in haptoglobin concentrations. In this study, CRP had 91% sensitivity and 61% specificity to predict mortality while haptoglobin had 52% sensitivity and 63% specificity (Kocaturk et al. 2010). Dogs naturally infected with Spirocerca lupi showed a sustained increase in CRP associated with neoplastic transformation of the oesophageal nodules which means a poor prognosis for outcome (Mukorera et al. 2011). Haptoglobin and CRP concentrations were 95 and 68% increased, respectively, in dogs with clinical signs and S. lupi ova on faecal specimens (Mylonakis et al. 2012). CRP showed 93% sensitivity for detecting Leishmania infantum symptomatic dogs and 82% sensitivity for detecting asymptomatic dogs (Martinez-Subiela et al. 2002). In experimental Leishmania infantum infection in dogs, CRP, serum amyloid A (SAA) and haptoglobin concentrations were higher before immunoglobulin G and immunoglobulin M increases and the appearance of clinical signs. During the entire infection period, haptoglobin remained increased and the treatment induced a significant decrease in all APPs (Martinez-Subiela et al. 2011). Dogs naturally infected with Babesia rossi and with clinical signs of acute babesiosis had higher CRP concentrations that decreased following treatment with imidocarb dipropionate (Rafaj et al. 2013). CRP was not associated with outcome in Babesia rossi infection in dogs but none of the survivors had on the admission day CRP concentrations less than 63·2 mg/L (Koster et al. 2009). In pyometra, SAA, CRP and haptoglobin were valid indicators of the inflammatory state of the uterus, making it possible to differentiate between open and closed cervix infections (Dabrowski et al. 2013) and to evaluate the severity of the inflammatory process especially with post-surgery complications (Dabrowski et al. 2009). In another study, 12 hours after standard soft-tissue surgeries (vasectomy, laparoscopic-assisted ovariohysterectomy and open approach ovariohysterectomy), CRP concentrations were significantly different and higher than prior to surgery ( Kjelgaard-Hansen et al. 2013). Investigation of APP in feline medicine has been relatively neglected and deserves to be further explored. The majority of investigations on feline APP have focused on pathophysiological conditions and have not generally examined time-series samples from cases. α 1 Acid glycoprotein (AGP), SAA and haptoglobin concentrations in groups of hospitalised cats, submitted to surgery and with induced inflammation were 7 to 11 times higher than in healthy cats but there was no significant change of CRP concentrations between the evaluated groups (Kajikawa et al. 1999). AGP and haptoglobin and also SAA and AGP showed strong correlations between sick cats (Kann et al. 2012), demonstrating that APPs should be evaluated as an auxiliary diagnostic tool in this species. AGP concentrations were significantly higher in tumour-bearing cats when compared to healthy cats, although no significant differences were found among the different groups of tumours (carcinomas, sarcomas and discrete round cell tumour) (Selting et al. 2000). In cats with lymphoma serum AGP concentrations did not provide useful information on response or survival, but the pretreatment AGP concentrations were significantly higher for cats with lymphoma than for healthy cats (Correa et al. 2001). A recent study with Mycoplasma haemofelis and “Candidatus Mycoplasma haemominutum” demonstrated that experimental infections associated or not with FIV infection in cats induced significant increases in APP concentrations and showed variable concentrations of haptoglobin, SAA and AGP according to the stage of infection (Korman et al. 2012). In cats with high antibody titers for chlamydiae, the SAA concentration was significantly higher when the bacteria were detected in conjunctival swab samples than when no chlamydiae were detected (Holst et al. 2011). For some time, AGP has been recognised as a biomarker of feline infectious peritonitis (FIP) (Duthie et al. 1997, Giordano et al. 2004, Paltrinieri et al. 2007) and has been reported to have 100% specificity and sensitivity in the diagnosis concordance for FIP (Giori et al. 2011). One of the few investigations of time-series monitoring of APP in cats has been carried out

Non-Invasive Measurement of Adrenocortical Activity in Blue-Fronted Parrots (Amazona aestiva, Linnaeus, 1758).

Parrots kept in zoos and private households often develop psychological and behavioural disorders. Despite knowing that such disorders have a multifactorial aetiology and that chronic stress is involved, little is known about their development mainly due to a poor understanding of the parrots’ physiology and the lack of validated methods to measure stress in these species. In birds, blood corticosterone concentrations provide information about adrenocortical activity. However, blood sampling techniques are difficult, highly invasive and inappropriate to investigate stressful situations and welfare conditions. Thus, a non-invasive method to measure steroid hormones is critically needed. Aiming to perform a physiological validation of a cortisone enzyme immunoassay (EIA) to measure glucocorticoid metabolites (GCM) in droppings of 24 Blue-fronted parrots (Amazona aestiva), two experiments were designed. During the experiments all droppings were collected at 3-h intervals. Initially, birds were sampled for 24 h (experiment 1) and one week later assigned to four different treatments (experiment 2): Control (undisturbed), Saline (0.2 mL of 0.9% NaCl IM), Dexamethasone (1 mg/kg IM) and Adrenocorticotropic hormone (ACTH; 25 IU IM). Treatments (always one week apart) were applied to all animals in a cross-over study design. A daily rhythm pattern in GCM excretion was detected but there were no sex differences (first experiment). Saline and dexamethasone treatments had no effect on GCM (not different from control concentrations). Following ACTH injection, GCM concentration increased about 13.1-fold (median) at the peak (after 3–9 h), and then dropped to pre-treatment concentrations. By a successful physiological validation, we demonstrated the suitability of the cortisone EIA to non-invasively monitor increased adrenocortical activity, and thus, stress in the Blue-fronted parrot. This method opens up new perspectives for investigating the connection between behavioural disorders and stress in this bird species, and could also help in their captive management.

Haematology of captive grey-brocket deer Mazama gouazoubira (Fischer, 1814) (Cervidae: Odocoileinae)

Reference intervals for haematologic and total plasma proteins profiles were determined for 11 physically restrained adult grey-brocket deer. Erythrocytic alterations, as discrete to moderate poikilocytosis, were observed in all animals. Red and white blood cells counts were determined by manual methodology. Obtained erythron and leukon values were slightly higher than previous reports. Statistical difference was not demonstrated between stags and hinds. No previous studies reported blood values for captive grey-brocket deer obtained by manual methods. Also, the authors suggest that future studies on deer haematology should be focused on technique evaluation and improvement, considering the unusual erythrocytic morphology.

Hematologic and Total Plasma Protein Values in Free-Living Red-tailed Amazon Parrot Nestlings (Amazona brasiliensis) in Paraná State, Brazil.

Abstract: The red-tailed Amazon parrot (Amazona brasiliensis) is an endangered psittacid species that is endemic in the south and southeast Brazilian Atlantic coastal region. Hematologic evaluation is important to monitor the health of these birds, and information about laboratory values for this species is scarce. Hematologic and total plasma protein profiles were determined for 33 free-living nestling parrots in Paraná state, Brazil. Parrots were temporarily removed from the nest and manually restrained to record body weight and collect blood samples. Mean body weight was <400 g in 13 birds (group 1) and >400 g in 20 birds (group 2). Significantly higher levels of mean corpuscular hemoglobin concentrations, white blood cell counts, monocytes, and basophils were observed in younger birds (group 1). A stress leukogram (high white blood cell and heterophil count) was found in all nestlings, suggesting stress induced by capture and restraint. Parameters obtained in this study will be essential to assess the physiologic and pathologic condition of wild parrots, to evaluate the effects of environmental changes on their health, and to contribute to conservation efforts of this endangered species.

Acute phase proteins as markers of infectious diseases in small animals.

Abstract During the acute phase response, there is an increased production and release of certain proteins known as acute phase proteins (APPs) which can be produced by hepatocytes and peripheral tissues such as C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin (Hp), alpha-1 acid glycoprotein (AGP). These proteins have been investigated as markers of various infectious diseases in small animals and the purpose of this review is to update the current knowledge about APPs in infectious diseases in dogs and cats. Tokom odgovora akutne faze, postoji poveć ana proizvodnja i oslobađanje određenih proteina, poznatih kao proteini akutne faze (APP), koje mogu proizvesti hepatociti i periferna tkiva. Proteini akutne faze su C-reaktivni protein (CRP), serumski amiloid A (SAA), haptoglobin (Hp), alfa-1 kiseli glikoprotein (AGP). Ovi proteini su ispitivani kao markeri raznih infektivnih oboljenja pasa i mačaka. Svrha ovog rada je da se ažuriraju savremena saznanja o APP tokom zaraznih bolesti pasa i mačaka

Proteínas de fase aguda em cadelas com neoplasia mamária

Acute phase proteins (APPs) are serum proteins whose concentrations change after infectious and inflammatory disease, and cancer. The aims of this study were to evaluate changes in APPs concentration and to correlate these findings with histological classification and WBC in female dogs with mammary tumors. APPs were studied in 45 female dogs with mammary tumor distributed in the following groups: benign (n=13), malignant without tumor ulceration (n=24), and malignant with tumor ulceration (n=8). SDS-polyacrylamide gel (SDS-PAGE) electrophoresis was used to measure APPs concentrations (albumin, ceruloplasmin, transferrin, haptoglobinHp, α-1-acid glycoprotein and α-1-antitrypsin) and ultrasensitive assay was used to evaluate serum C-reactive protein (CRP). Patients with malignant mammary neoplasia plus ulceration had significant increase of CRP and Hp, and had decreased levels of albumin (P<0.05, One-Way ANOVA and Dunn Test). Positive correlation among APPs and inflammatory leukocytosis were observed (P=0.002, Fisher test). No correlation was observed between APPs and histological subtype. In conclusion, combined changes of CRP, Hp and albumin may be used as a prediagnostic tool and prognosis in dogs with mammary tumors.

Efeito do sítio de venopunção nos parâmetros hematológicos em tigre-d'água-americano, Trachemys scripta elegans

The aim of the present study was to compare two different blood collection sites of 24 red-eared sliders (Trachemys scripta elegans) from a commercial breeder, situated in Antonina, Parana, Brasil. The animals were physically restrained and paired blood samples were collected from the dorsal coccygeal vein and the occipital sinus. The samples were collected in syringes containing heparin. After collection the samples were identified and refrigerated to posterior hematological analysis. Red blood cell and white blood count were performed using a hemocytometer. The packed cell volume and hemoglobin were determined by the microhematocrit method and cyanmethemoglobin method, respectively. Total plasma protein was determined by refractometry and differential leukocyte count was performed by the Shilling technique. There was a significant difference in the total white blood cell count and on the total plasma protein value, and on both cases the results found on the samples from the dorsal coccygeal vein were lower. The difference found on the white blood cell count was probably due to the lymph contamination of the samples, which also justifies the lower values of total plasma protein. In conclusion, the occipital sinus is more recommended blood collection site than the dorsal coccygeal vein.

Mel e açúcar mascavo na cicatrização de feridas

The study aimed to compare the effectiveness of honey and brown sugar in wounds healing induced in the skin of guinea pigs, to evaluate the healing time, microscopic and macroscopic healing variations, wound contraction and contamination degree. Fifteen guinea pigs, Abyssinia breed, males, six months of age and weighing between 500 to 850 grams, were divided in three groups with five animals each. Three wounds were promoted in each animal, surgical, with 1.5cm² and 1.5cm between them, in the dorsal region. The F1 was treated, daily, with honey, F2 with saline solution 0,9% (control) and F3 with brown sugar. Euthanasia was performed in Group 1 on the seventh day after the surgery, Group 2 on the fourteenth day and the Group 3 on the twenty-first day after the surgery and the biopsies sent to histopathological evaluation. Wounds swabs were made in all groups on the third and seventh days after the surgery and the samples sent for culture and bacterial isolation. The wounds treated with honey healed faster than treated with brown sugar.

Changes in biochemical analytes in calves infected by nematode parasites in field conditions.

Parasitic infections caused by nematodes are a major problem in bovines that resulting in losses in animal health and production. Thus, the aim of this study was to evaluate alterations in selected serum biochemical analytes in calves naturally infected with gastrointestinal (GI) and pulmonary nematodes without clinical signs. For this, samples of feces and blood of 86 calves were collected. Fecal egg counts (FEC) were determined using the modified McMaster technique with a sensitivity of 50 eggs per gram of feces (EPG). Positive nematode FEC was processed for coproculture using pooled samples to identify Strongylidae infective larvae (L3). First stage-larvae (L1) of Dictyocaulus viviparous were identified by a modified Baermann method. The biochemical analytes determined were: acute phase proteins such as haptoglobin and paraoxonase type 1; the enzymes acetylcholinesterase; butyrylcholinesterase; the lipid profile (triglycerides and total, HDL, and LDL-cholesterol); serum iron profile (iron and unsaturated iron-binding capacity); total protein and albumin; pancreatic profile (amylase and lipase); and minerals (phosphorus and calcium). The calves were divided into four groups according to the results of EPG and the modified Baermann method. Group 1: healthy control animals (n=16); Group 2: calves with only GI parasites (n=51): This group was sub-divided into sub-groups according to the EPG threshold: 2a-GI parasites with low EPG (n=23), and 2b-GI parasites with high EPG (n=28). Group 3: animals with only lungworms (n=5), and Group 4: calves with lung+GI parasites (n=14). The more prevalent genera in all coprocultures were: Cooperia spp., Haemonchus spp., Oesophagostomum spp., and Trichostrongylus spp. The nonparametric Kruskal-Wallis test was used to compare the groups and Dunns post-test was used for multiple comparisons as the data was not normally distributed (P<0.05). The haptoglobin concentration increased in calves with GI and pulmonary parasites. A significant increase in acetylcholinesterase was observed in calves infected with lungworms. Cholesterol, triglycerides, HDL, and LDL concentrations decreased but lipase concentration increased in calves with GI parasites. Therefore, this paper provides an overview of the biochemical effects produced by nematode parasites in calves in field conditions. These findings in calves without any evident clinical signs of disease could provide an indication of GI parasites and lungworm infection, especially in an endemic area for these parasites.