Automated flow cytometric cell count and differentiation of canine cerebrospinal fluid cells using the ADVIA 2120

Background: Microscopic cell counts in cerebrospinal fluid (CSF) are time-consuming and prone to imprecision. The recently introduced automated hematology analyzer ADVIA 2120 offers an automated cell count and differential for CSF in the veterinary software mode based on laser light scatter and absorbance measurements. Objectives: The purpose of this study was to evaluate the precision, linearity, and accuracy of the ADVIA 2120 CSF assay. Methods: Sixty-seven CSF samples were analyzed on the ADVIA 2120 and total nucleated cell counts (TNCC) and RBC counts were compared with the hemocytometer results. In 21 samples with TNCC >5/muL, ADVIA 2120 results were compared with 100-300 cell manual differentials performed on cytocentrifuged preparations. Statistical analysis included Spearman's rank correlation, Passing-Bablok regression, and Bland-Altman analysis. Results: Repeatability (intra-assay) coefficients of variation (CVs) ranged from 4.19% to 25.94%. Interassay CVs ranged from 2.56% to 28.67%. Accurate results within 30% were achieved for TNCC up to 4000/muL. Except for low TNCC, deviation from the expected value was higher (TNCC of 8/muL instead of 4/muL). The following correlation coefficients (r) and biases were achieved compared with the reference method: r=.90 and bias 2.3/muL for TNCC; r=.88 and bias 32.0/muL for RBC counts; r=.86 and bias +/-13.4% for mononuclear and polymorphonuclear cell percentages; r=.88 and bias -6.1% for lymphocyte percentage; r=.56 and bias 19.4% for monocyte percentage; and r=.75 and bias -9.7% for neutrophil percentage. Conclusion: Our results demonstrated that the automated ADVIA 2120 CSF assay generally compares well with reference methods although there are some limitations for the automated monocyte count and for samples with only mild pleocytosis.     

Comparative clinical study of canine and feline total blood cell count results with seven in‐clinic and two commercial laboratory hematology analyzers

Background: A CBC is an integral part of the assessment of health and disease in companion animals. While in the past newer technologies for CBC analysis were limited to large clinical pathology laboratories, several smaller and affordable automated hematology analyzers have been developed for in‐clinic use. Objectives: The purpose of this study was to compare CBC results generated by 7 in‐clinic laser‐ and impedance‐based hematology instruments and 2 commercial laboratory analyzers. Methods: Over a 3‐month period, fresh EDTA‐anticoagulated blood samples from healthy and diseased dogs (n=260) and cats (n=110) were analyzed on the LaserCyte, ForCyte, MS45, Heska CBC, Scil Vet ABC, VetScan HMT, QBC Vet Autoread, CELL‐DYN 3500, and ADVIA 120 analyzers. Results were compared by regression correlation (linear, Deming, Passing‐Bablok) and Bland–Altman bias plots using the ADVIA as the criterion standard for all analytes except HCT, which was compared with manual PCV. Precision, linearity, and carryover also were evaluated. Results: For most analytes, the in‐clinic analyzers and the CELL‐DYN performed similarly and correlated well with the ADVIA. The biases ranged from ?0.6 to 2.4 × 10⁹/L for WBC count, 0 to 0.9 × 10¹²/L for RBC count, ?1.5 to 0.7 g/dL for hemoglobin concentration, ?4.3 to 8.3 fL for MCV, and ?69.3 to 77.2 × 10⁹/L for platelet count. Compared with PCV, the HCT on most analyzers had a bias from 0.1% to 7.2%. Canine reticulocyte counts on the LaserCyte and ForCyte correlated but had a negative bias compared with those on the ADVIA. Precision, linearity, and carryover results were excellent for most analyzers. Conclusions: Total WBC and RBC counts were acceptable on all in‐clinic hematology instruments studied, with limitations for some RBC parameters and platelet counts. Together with evaluation of a blood film, these in‐clinic instruments can provide useful information on canine and feline patients in veterinary practices.     

Neuropathological survey of fallen stock: active surveillance reveals high prevalence of encephalitic listeriosis in small ruminants

This paper describes the prevalence of brain lesions in the Swiss fallen stock population of small ruminants. 3075 whole brains (75% sheep, 25% goats) were collected as part of a year-long active survey of transmissible spongiform encephalopathies (TSEs) in small ruminants conducted by the Swiss authorities between July 2004 and July 2005. All fallen stock brains were systematically examined by histopathology to obtain reliable data on histologically identifiable brain lesions. Lesions were found in an unexpectedly high number of animals (8.1% of all examined brains). A wide spectrum of diseases was detected showing that this approach provides an excellent opportunity to screen for the prevalence of neurological diseases. Encephalitic listeriosis was by far the most frequent cause of CNS lesions in both species and its prevalence was unexpectedly high when compared to notified confirmed cases. In conclusion, the prevalence of listeriosis as estimated by passive surveillance based on the notification of clinical suspects has been underestimated in the past.     

Morphological indices for canine spermatozoa based on the World Health Organization laboratory manual for human semen

Attempting to contribute to the development of a more objective morphological evaluation of dog spermatozoa, in this study the indices of multiple sperm defects (multiple abnormalities index [MAI]; teratozoospermic index [TZI]; sperm deformity index [SDI]) were calculated following the World Health Organization (WHO) guidelines. In Experiment I, the concordance of MAI, TZI and SDI with the proportions of morphologically normal spermatozoa (MNS) was evaluated in fresh ejaculated spermatozoa (dogs = 47). In Experiment II, the potential role of indices as prognostic values was assessed in spermatozoa of different origin and treatment (fresh ejaculated: n = 6; fresh epididymal: n = 6; frozen‐thawed ejaculated: n = 6) by their correlation with different semen parameters (motility, membrane integrity and acrosome status) and with an in vitro sperm function test. Samples with different proportions of MNS showed different values of SDI, the index that better represented the decline of sperm morphology in both fresh and frozen‐thawed samples (Exp. I and II; p < 0.05). No correlations between indices and semen parameters were observed (Exp. II), but when samples were evaluated collectively, negative correlations (SDI‐motility, p = 0.01; SDI‐acrosome integrity, p = 0.002) were found. Including all the defects of each spermatozoon, SDI might be a useful index during morphological analysis and better discriminates the increase in multiple defects. A more objective morphological evaluation for dog spermatozoa was achieved by the WHO method, and in vitro tests allowed to elucidate the validity of SDI as prognostic indicator of in vitro fertilizing potential.     

Quality of Wine Grape Cultivars Grown in Czech Republic

This research aims to make an analysis of significant parameters in wine grape from Czech Republic. It was evaluated pH, content of sugar, ascorbic acid, titratable acidity, total phenolic content and texture properties. For analyzes 5 samples of wine grape were used, harvested during the three periods. The results show that during the ripening of wine grapes there were changes of selected qualitative parameters, particularly of total polyphenols, sugar and some varieties of wine grape also ascorbic acid (significantly in the variety ’Grüner Veltliner’, ’Chardonnay’, ’Pinot Blanc’). Pronounced changes qualitative parameters were among wine grapes harvested the 1st and 2nd seasons. Changes in textural properties also occurred during the harvest and for most varieties between 2nd and 3rd period of harvest. The PCA grouped chemical parameters of wine grape in independent sets and can be used as assessment tool of wine cultivars.     

Ultrasonographic differentiation between portal venous and parenchymal gas may be important for the prognosis of canine and feline hepatic emphysema: 37 cases

The aim of this retrospective, cross‐sectional, study was to evaluate clinical findings and outcomes for different ultrasonographic patterns of hepatic emphysema in dogs and cats. Dogs and cats with an ultrasonographic diagnosis of hepatic emphysema and a known outcome, from January 2010 to January 2018, were enrolled. The following data were recorded from medical and ultrasonographic records: ultrasonographic patterns of hepatic emphysema (parenchymal, portal venous, biliary), clinical signs, laboratory findings, and outcomes (favorable, poor). A total of 33 dogs and four cats met the inclusion criteria. Among these, 23 cases were classified as hepatic portal venous gas, 10 as parenchymal emphysema, and four as biliary emphysema. Clinical diagnosis categories were as follows: infection/sepsis (9), gastro‐intestinal disease (9), iatrogenic (9), trauma (5), and liver neoplasia (5). An increase in serum liver enzymes was significantly associated with parenchymal emphysema (P = .03). Other clinical and laboratory findings were not associated with the type of hepatic emphysema. Hepatic portal venous gas was mostly transient in patients with ultrasonographic follow‐up. The overall mortality was 40.5%. A significant difference was found between mortality by portal venous gas (21.7%) and mortality by parenchymal emphysema (90%) (P = .003). In conclusion, the ultrasonographic differentiation of hepatic emphysema between hepatic portal venous gas and parenchymal emphysema may be important for the prognosis of hepatic emphysema. The presence of parenchymal emphysema may be a poor prognostic indicator, while hepatic portal venous gas may be more benign. However, ultrasound findings should be carefully evaluated in the context of clinical findings.     

Screening of Uruguayan plants for deterrent activity against insects

We evaluated the anti-insectan activity of extracts from different vegetative parts of ten plant species native to Uruguay. The selected plants belong to five families: Bignoniaceae: Clytostoma callistegioides, Dolichandra cynanchoides, Macfadyena unguis-cati; Sapindaceae: Dodonaea viscosa, Allophylus edulis, Serjania meridionalis; Lamiaceae: Salvia procurrens, Salvia guaranitica; Solanaceae: Lycium cestroides; and Phytolaccaceae: Phytolacca dioica. The extracts were evaluated in independent bioassays against four insect pests and one beneficial insect. Aphid settling inhibition was evaluated with a grass specialist, Rhopalosiphum padi, and a feeding generalist, Myzus persicae (both Hemiptera: Aphididae). Antifeedant activity was tested with adults of the specialist Epilachna paenulata (Coleoptera: Coccinellidae) and larvae of the generalist Spodoptera littoralis (Lepidoptera: Noctuidae). Finally, contact toxicity was assessed with honey bees, Apis mellifera (Hymenoptera: Apidae). Strong settling inhibition (SI) activity (expressed as %SI, where 100% means complete inhibition by the extract) was found only for the twig extracts of A. edulis (Sapindaceae) against M. persicae (%SI = 77 ± 4). Antifeedant activity (expressed as % of feeding reduction (FR), where 100% means no consumption on extract-treated diet) against E. paenulata was significant for the leaf extracts of L. cestroides (Solanaceae) (%FR = 100 ± 0) as well as of all Bignoniaceae and Sapindaceae species. No extracts were active against S. littoralis larvae, and most of them were innocuous to honey bees, with the exception of L. cestroides and S. meridionalis leaf extracts.     

Emissions of ammonia,nitrous oxide and carbon dioxide from urban gardens in Niamey,Niger

Urban and peri-urban agriculture (UPA) contributes significantly to meet increasing food demands of the rapidly growing urban population in West Africa. The intensive vegetable cultivation in UPA gardens with its high nutrient inputs is often reported to operate at large surpluses of nutrients and presumably high turnover rates of organic matter (OM) and nitrogen (N) losses via emanation and leaching. Many of these claims are lacking solid data which would allow suggesting mitigation strategies. Therefore, this study aimed at quantifying gaseous emissions of ammonia (NH₃), nitrous oxide (N₂O), and carbon dioxide (CO₂) in three representative urban gardens of Niamey, Niger using a closed chamber gas monitoring system. Mean annual N emissions (NH₃-N and N₂O-N) in two gardens using river water for irrigation reached 53 and 48 kg N ha^?1 yr^?1, respectively, while 25 and 20 Mg C ha^?1 yr^?1 was lost as CO₂-C. In the garden irrigated with sewage water from the city's main wadi, N₂O was the main contributor to N losses (68%) which together with NH₃ reached 92 kg N ha^?1 yr^?1, while CO₂-C emissions amounted to 26 Mg ha^?1 yr^?1. Our data indicate that 28% of the total gaseous C emissions and 30–40% of the N emissions occur during the hot dry season from March to May and another 20–25% and 10–20% during the early rainy season from June to July. Especially during these periods more effective nutrient management strategies in UPA vegetable gardens should be applied to increase the nutrient use efficiency in UPA vegetable gardens.     

Phosphorus forms and enzymatic hydrolyzability of organic phosphorus in soils after 30 years of organic and conventional farming

Lower P‐input levels in organic than conventional farming can decrease soil total and available P, which can potentially be resupplied from soil organic P. We studied the effect of 30 y of conventional and organic farming on soil P forms, focussing especially on organic P. Soil samples (0–20 cm) were taken in a field experiment with a nonfertilized control, two organic systems receiving P inputs as animal manure, and two conventional systems receiving only mineral P or mineral P and manure. Soils were analyzed for total, inorganic, organic, and microbial P, by sequential P fractionation and by enzyme additions to alkaline soil extracts. Samples taken prior to starting the experiment were also analyzed. Average annual P balances ranged from –20 to +5 kg ha^–1. For systems with a negative balance, labile and moderately labile inorganic P fractions decreased, while organic and stable inorganic P fractions were hardly affected. Similar quantities and proportions of organic P extracted with NaOH‐EDTA were hydrolyzed in all soils after addition of an acid phosphatase, a nuclease, and a phytase, and enzyme‐stable organic P was also similar among soils. Thus, neither sequential fractionation nor enzyme addition to alkaline soil extracts showed an effect of the type of applied P (manure vs. mineral) on organic P, suggesting that organic P from manure has largely been mineralized. Thus far, we have no indication that the greater microbial activity of the organic systems resulted in a use of stable P forms.