Assessing failure of transfer of passive immunity by gamma-glutamyl-transferase activity and serum refractometry in holstein-friesian calves affected by neonatal diarrhea.

Hardly published data are available to diagnose the failure of transfer of passive immunity (FTPI) in calves affected by neonatal calf diarrhea (NCD). This study evaluated the diagnostic performance and differences among optical serum total protein (STP) concentration and gamma-glutamyl-transferase (GGT) activity for assessing FTPI in diarrheic Holstein Friesian calves. 72 diarrheic and 19 healthy Holstein Friesian calves aged 1 to 10 days were enrolled. Each calf underwent a complete clinical examination and dehydration assessment. The effect of dehydration status and age on the correlation between the two methods under study (STP and GGT) and the gold standard (Immunoglobulin G [IgG] measured with RID) was investigated with Spearman's correlation index R for ranks. Serum total protein concentration and GGT activity were analyzed by receiver operating characteristic (ROC) curve analysis to identify the optimal cut-off point to distinguish between diarrheic calves with or without FTPI, also considering the effects of dehydration and age. The results show that GGT activity was affected by the age of calves, while STP was influenced by dehydration. The cut-offs to distinguish calves with IgG < 10 g/L were < 52 g/L of STP in normohydrated calves, < 58 g/L of STP in dehydrated calves, and < 124 IU/L of GGT in calves with aged between 3 and 10 days. In nondehydrated diarrheic calves, the STP refractometer showed better diagnostic accuracy.In dehydrated calves aged between 3 and 10 days, however, its accuracy drops, and it is advisable to use GGT activity.

Comparison of two injectable anaesthetic protocols in Egyptian fruit bats (Rousettus aegyptiacus) undergoing gonadectomy.

Egyptian fruit bats have gained increasing interest being a natural reservoir for emerging zoonotic viruses. Anaesthesia is often required to allow safe handling of bats. We aimed to compare the sedative and cardiopulmonary effects of two balanced anaesthetic protocols in bats undergoing gonadectomy. Group DK (n = 10) received intramuscular dexmedetomidine (40 µg/kg) and ketamine (7 mg/kg), whereas group DBM (n = 10) received intramuscular dexmedetomidine (40 µg/kg), butorphanol (0.3 mg/kg) and midazolam (0.3 mg/kg). Induction time and cardiopulmonary parameters were recorded. If anaesthetic plan was inadequate, isoflurane was titrated-to-effect. At the end of surgery venous blood gas analysis was performed and atipamezole or atipamezole-flumazenil was administered for timed and scored recovery. In DBM group heart rate and peripheral oxygen saturation were significantly higher (p = 0.001; p = 0.003 respectively), while respiratory rate was significantly lower (p = 0.001). All bats required isoflurane supplementation with no significant differences between groups. Induction and recovery times showed no significant differences. In group DK a better recovery was scored (p = 0.034). Sodium and chloride were significantly higher in DBM group (p = 0.001; p = 0.002 respectively). Both anaesthetic protocols were comparable and can be recommended for minor procedures in bats.

Clinical and Clinical Pathological Presentation of 310 Dogs Affected by Lymphoma with Aberrant Antigen Expression Identified via Flow Cytometry.

Phenotypic aberrancies have been reported occasionally in canine lymphomas. Here, we retrospectively collected 310 canine lymphomas with an aberrant phenotype detected via flow cytometry and describe their clinical and clinical pathological features at diagnosis. There were 152 T-cell lymphomas not otherwise specified (T-NOS), 101 T-zone lymphomas (TZL), 54 B-cell lymphomas, and 3 cases with two suspected concurrent neoplastic populations. The most represented aberrancies were: CD5-, CD4-CD8-, and CD3- in T-NOS lymphomas, CD21+, CD4-CD8-, and CD3- in TZLs, and CD34+, CD44-, and CD5+ in B-cell lymphomas. Among T-cell lymphomas, the aberrant expression of CD21 was significantly more frequent in TZL and the loss of CD5 and CD44 in T-NOS. More than 75% of dogs were purebred; males outnumbered females; the mean age at diagnosis was 8-10 years, depending on lymphoma subtype. A few dogs were symptomatic at the time of diagnosis, and 30% had peripheral blood abnormalities, in line with what is already reported for the general population of dogs with lymphoma. Further studies are needed to assess the pathogenetic mechanisms underlying each specific antigen aberrancy, as well as the diagnostic and prognostic role.

Preliminary study on the effect of season on urinary analytes in healthy Italian dairy cows.

BACKGROUND: Season is known to affect serum analyte concentrations in dairy cows, and the same can be hypothesized for urinary analytes, but information in this regard is lacking. OBJECTIVES: The aim of the study was to assess the effect of seasonality on urinary variables in healthy dairy cows. METHODS: Twelve Italian Holstein cows were randomly selected from a local herd, and urine specimens were collected once in each season of the year. For each cow, the lactation stage at each sampling time was also registered (lactation vs dry period), and physical examination and hematology and serum biochemistry measurements were used to assess health status. Three cows were voluntarily culled from the herd during the year; therefore, nine cows were included. Concentrations of urinary analytes recorded in each season for each cow were compared. RESULTS: Seasonality affected urinary protein-to-creatinine ratios (P = 0.012) with lower median values in summer compared with spring (P = 0.020) and autumn (P = 0.004); differences were also found in urinary sodium-to-urinary creatinine ratio (P = 0.009), with lower medians in summer compared with spring (P = 0.003) and autumn (P = 0.020). The direct consequences of higher summer temperatures and the acclimation strategies needed to adapt to this environment could explain the changes in urinary analytes that were recorded in the current study; in fact, decreased food and water consumption tend to increase urinary creatinine concentrations and decrease renal excretion of proteins and electrolytes. CONCLUSIONS: The present results suggest that seasonality can affect urinary variables of healthy dairy cows.

Analytical variability and uncertainty in canine leukocyte ratios obtained with manual counts.

BACKGROUND: This study aimed to determine the analytical imprecision of calculated neutrophil to lymphocyte (NLR) and lymphocyte to monocyte ratios generated from manual differential white blood cell (WBC) counts in peripheral blood smears, and to describe how to report the uncertainty around a single WBC ratio result. No information on the analytical imprecision of WBC ratios in dogs is available. METHODS: Coefficient of variations (CVs) of paired readings of one operator on 105 smears (intraoperator variability) and of three operators on 301 smears (interoperator variability) were calculated. The interoperator agreement was examined with the Fleiss' kappa coefficient (κ). Observed total errors (TEos), expanded measurement of uncertainty (EMU) and reporting intervals (RIs) were also calculated. RESULTS: Median CVs ranged from 3.14 to 28.28 (intraoperator) and from 5.39 to 53.85 (interoperator). No agreement among operators was found around the cut-offs. TEos were higher than allowable total errors in 32%-88% of smears. EMU ranged from 0.10 to 1.13. According to the RI, the calculated WBC ratios should be rounded to the nearest 10. CONCLUSION: WBC ratios should be interpreted cautiously in dogs. The EMU should be reported to make the clinician aware of the uncertainty of these parameters. For example, an NLR result of 17 is needed to have high confidence that the result is above a cut-off of 6.

Analytical Validation of a New Immunoenzymatic Method for the Measurement of Feline Parathyroid Hormone in Cats with Chronic Kidney Disease.

The determination of parathyroid hormone (PTH) in cats could be of clinical utility in many metabolic disorders, such as renal diseases, hypercalcemia, or nutritional imbalances. However, the available methods for the measurement of feline PTH are limited, not widely available, and need radioimmunoassays. The aim of this study was to perform the analytical validation of a new immunoenzymatic method for the measurement of feline PTH. Thirty-eight cats affected with chronic kidney disease (CKD) were included. PTH was measured using a two-site immunoenzymatic method validated in humans and dogs (ST AIA-PACK® Intact PTH, Tosoh Bioscience, Tessenderlo, Belgium). The analytical validation provided the evaluation of precision (intra-assay and inter-assay), accuracy (linearity under dilution (LUD) and spike recovery test (SRT)), and the storage stability of serum samples at 20 °C, 4 °C, and -20 °C. The method showed good precision (intra-assay CVs (coefficient of variations) 3.19-9.61%; inter-assay CVs 9.26-15.28%). In both the intra- and inter-assays, the highest imprecision was found with the low concentration pool (9.61% and 15.28%) and accuracy (LUD and SRT r2 = 0.99, p < 0.001), while the stability was optimal up until 7 days at -20 °C (-7.7%). The method was successfully validated in cats, allowing its future use in diagnostic procedures.

Haematological, serum biochemical and electrophoretic data on healthy captive Egyptian fruit bats (Rousettus aegyptiacus).

Bats play a key role as reservoir hosts of many emerging viral diseases with zoonotic potential. However, little is known about the laboratory reference intervals (RIs) of bats, especially Egyptian fruit bats (Rousettus aegyptiacus). The aim of this study was to obtain haematological, biochemical and electrophoretic RIs from captive fruit bats. Blood was collected from 21 R. aegyptiacus (11 females and 10 males). Complete blood cell count was performed using an impedance cell counter followed by the morphologic analysis of blood smears. Clinical biochemistry was performed with an automated spectrophotometer and agarose gel electrophoresis was carried out with an automated instrument. Reference intervals were determined using the Reference Value Advisor V2.1, following the American Society for Veterinary Clinical Pathology guidelines. Possible differences related to sex or sexual maturity were also investigated. The RIs for most of the analytes investigated were similar to those of other types of bats and other mammalian species. Haematology revealed mild polychromasia and slightly lower haematocrit, haemoglobin, leukocyte and lymphocyte counts compared to other bats. Glucose levels varied possibly due to stress, the anaesthetic protocol and fasting time. Creatine kinase was higher, while triglycerides were lower compared with domestic mammals and other bats. No sex- or age-related differences were found. Serum protein electrophoresis showed five fractions (albumin, α-, ß1-, ß2- and γ-globulins). The values recorded in this study could be helpful as a reference biological dataset to monitor the health status of wild and captive R. aegyptiacus and, possibly, of other Chiroptera.

Analytical and Clinical Validation of a New Immunoenzymatic Method for the Measurement of Canine Parathyroid Hormone.

Renal hyperparathyroidism (RHPT) is one of the main complications in dogs affected with Chronic Kidney Disease (CKD). The measurement of serum parathyroid hormone (PTH) could be of clinical utility for the disease's treatment and follow-up; however, PTH is not routinely determined due to limited available methods, often not fully validated in dogs. The aims of this study were the analytical validation of an immunoenzymatic method for the measurement of PTH in canine serum and the analysis of preliminary association of the obtained results with renal function. Twenty-six samples obtained from dogs healthy or affected with CKD were analysed. PTH was measured using a two-site immunoenzymometric human assay (ST AIA-PACK® Intact PTH, Tosoh Bioscience). The analytical validation protocol evaluated the assay precision and accuracy. Also, the PTH's storage stability at 20 °C, 4 °C and -20 °C was assessed. Clinical validation was performed by comparing PTH values with creatinine, phosphorus and International Renal Interest Society (IRIS) stage. The method showed optimal precision and accuracy, whereas stability was adequate up to 4 h at 20 °C, 24 h at 4 °C and 6 months at -20 °C. PTH was positively associated with creatinine, phosphorus and IRIS stage. The investigated method was thus successfully validated in dogs, allowing its use for clinical purpose.

Reference intervals for hematological and biochemical parameters, acute phase proteins and markers of oxidation in Holstein dairy cows around 3 and 30days after calving.

The establishment of specific reference intervals (RIs) is advisable when different metabolic conditions in the patient population are present or when existing RIs are not suitable. Holstein dairy cows, at the start of lactation, experience a negative energy balance and an altered immune function that can lead to different levels of blood parameters compared with cows at peak lactation. The aim of this study was the determination of RIs for Holstein cows at 3±1 and 30±3days in milk. To this aim, 145 cows, from 4 herds, were sampled. A wide panel of hematological and biochemical analytes was determined, as well as the measurement of markers of inflammation/oxidation. RIs were generated following the American Society of Veterinary Clinical Pathology (ASVCP) guidelines and the effects of lactation period, parity, herd and day of sampling were determined. Data from 39 out of 52 analytes were significantly different according to lactation period. Data of 13 out of 52 analytes were not significantly different between the two groups. Some differences were also recorded between herds and parity groups but the adoption of specific RIs for these subgroups, however, has practical limitations. In conclusion the use of RIs specific for the lactation period is highly justified from both a statistical and a biological point of view. Pre-analytical factors associated with parity or management need to be considered in the evaluation of results of some analytes.

Colorimetric and electrophoretic evaluation of lipoprotein fractions in healthy neonatal calves: Comparison with results from adult cows and from calves with inflammatory conditions.

High density lipoproteins (HDLs) are pivotal in innate immunity and decrease in serum during inflammation. Several studies have been done about lipoprotein changes in transition cows but little is known about their changes in newborn calves. The aim of this study is to provide information about HDLs in newborn calves, by defining the possible age-related changes in healthy calves compared with adults and by assessing the possible differences in calves with inflammation. Lipoprotein electrophoretic separation (reported as percentages) and colorimetric measurement of HDL (HDL-C) were performed on healthy cows and calves in order to identify possible differences in the lipoprotein profile due to the age. Then, age-matched calves with inflammatory conditions were also evaluated. Results showed that in calves HDL% and VLDL% were lower (mean values±SD: 77.6%±8.6% and 2.6%±2.5%, respectively) and LDL% was higher (19.7%±7.4%) than in adults (89.0%±3.9%; 5.2±2.1% and 5.8%±3.1%, respectively). Sick calves revealed a decrease of both HDL% (mean values ± SD: 61.0%±22.1%) and HDL-C (22.8±11.6mg/dL) and an increase of VLDL% (12.1% ±13.1%) compared with controls (77.6%±8.6%; 41.5±11.2mg/dL and 2.6%±2.5%, respectively). Paraoxonase-1 activity, influenced by inflammation and oxidation, was measured, and it appeared correlated with HDL% and HDL-C in sick calves. In conclusion, this study revealed that HDLs concentration in healthy calves is lower than in adults, and further decreases in calves with inflammation, likely due to oxidation.

Fluctuation of neutrophil counts around parturition in Holstein dairy cows with and without retained placenta.

Retained placenta (RP) is often diagnosed in high-yielding dairy cows and can negatively affect reproductive performances. The objective of the present study was to investigate the hematological and biochemical profile of cows with RP before and immediately after parturition, with particular emphasis on neutrophil counts, since a previous study demonstrated the presence of peripheral neutropenia in dairy cows with RP sampled a few days after parturition. Results from 12 Holstein cows affected by RP and from 17 clinically healthy controls sampled one week pre-partum, within 12h after calving and between 48 and 72h after parturition were compared between groups and over time. Compared with controls, cows with RP had lower lymphocyte counts before parturition, lower leukocyte and neutrophil counts at parturition, lower monocyte counts at all times, and higher ß-hydroxybutyrate before and after parturition. Erythroid and biochemical parameters were similar over time in both groups, whereas RP cows did not show the increase of neutrophil counts that occurs in controls at parturition. Hence, the finding of a lower neutrophil count in a routinely hemogram performed at parturition could be used as an alarm signal suggesting to monitor the affected animals. Moreover, although the underlying pathogenetic mechanism should be better investigated, the present study describes for the first time the association between altered blood leukocyte concentrations at parturition in RP compared to control cows.

Peripheral blood lymphocyte/monocyte ratio as a useful prognostic factor in dogs with diffuse large B-cell lymphoma receiving chemoimmunotherapy.

Diffuse large B-cell lymphoma (DLBCL) is the most frequent canine lymphoid neoplasm. Despite treatment, the majority of dogs with DLBCL experience tumour relapse and consequently die, so practical models to characterise dogs with a poor prognosis are needed. This study examined whether the lymphocyte/monocyte ratio (LMR) can predict outcome in dogs with newly diagnosed DLBCL with regard to time-to-progression (TTP) and lymphoma specific survival (LSS). A retrospective study analysed the prognostic significance of LMR obtained at diagnosis by flow cytometry (based on morphological properties and CD45 expression) in 51 dogs that underwent complete staging and received the same treatment, comprising multi-agent chemotherapy and administration of an autologous vaccine. Dogs with an LMR ≤ 1.2 (30% of all cases) were found to have significantly shorter TTP and LSS, and it was concluded that LMR was a useful independent prognostic indicator with biological relevance in dogs with DLBCL treated with chemoimmunotherapy.

Early post-partum hematological changes in Holstein dairy cows with retained placenta.

Retained placenta (RP) occurs frequently in dairy cattle but little is known about the pathogenic or prognostic role of the hematological changes in this disease. This retrospective study was designed to investigate the hematological changes associated with RP in the immediate post-partum period and to assess whether these changes are associated with an acute phase reaction. Data concerning hematology, acute phase proteins, markers of inflammation and serum biochemistry performed on cows at 3±1 days in milk (DIM) from two intensive farms were extracted from the database of the ProZoo project, a research project aimed to investigate the relationship between genomic traits and bovine health and production. After application of restrictive inclusion criteria, data from 45 cows, 22 with RP and 23 controls, were statistically compared. RBC count, d-ROMs concentration, and AST activity were significantly higher in the RP group than controls. Conversely, neutrophils, thiol groups, and serum zinc concentration were significantly lower in the RP group than controls. In conclusion, although retained placenta has to be considered as a syndrome with multifactorial causes, neutropenia may be a co-factor involved in its pathogenesis. Further studies are needed to clarify whether neutropenia acts as a contributor in the pathogenesis of RP or if it is a very early consequence of the syndrome, preceding any other inflammatory changes in blood.