Prior and concomitant dehydroepiandrosterone treatment affects immunologic response of cultured macrophages infected with Trypanosoma cruzi in vitro?

DHEA, a steroid hormone synthesized from cholesterol by cells of the adrenal cortex, plays an essential role in enhancing the host's resistance to different experimental infections. Receptors for this hormone can be found in distinct immune cells (especially macrophages) that are known to be the first line defense against Trypanosoma cruzi infection. These cells operate through an indirect pathway releasing nitric oxide (NO) and cytokines such TNF-α and IL-12 which in turn trigger an enhancement of natural killer cells and lymphocytes which finally secrete pro and anti-inflammatory cytokines. The effects of pre- and post-infection DHEA treatment on production of IL-12, TNFα and NO were evaluated. T. cruzi infected macrophages post treated with DHEA displayed enhanced concentrations of TNF-α, IL-12 and NO. Probably, the mechanisms that induced the production of cytokines by infected cells are more efficient when the immune system has been stimulated first by parasite invasion, suggesting that the protective role of DHEA is greater when administered post infection.     

Neutrophilic myeloperoxidase index and mean light absorbance in neonatal septic and nonseptic foals

Background: Two neutrophilic indices reported by the ADVIA 120 Hematology Analyzer, neutrophilic myeloperoxidase index (MPXI), and mean light absorbance (neutrophil X mean [NXM]) have been proposed as indicators of systemic inflammatory disease in horses and of neutrophil activation in coronary ischemic syndromes in people. Objective: The aim of this study was to evaluate NXM and MPXI in healthy, sick nonseptic, and sick septic foals to determine whether conditions likely associated with neutrophil activation result in decreases in these variables. Methods: In this retrospective study, CBC data from 61 neonatal foals presented to the Equine Teaching Hospital of Barcelona were evaluated for correlations between MPXI, NXM, percentage of large unstained cells, neutrophil count, and percentage of band neutrophils. Results obtained in septic (n=32), sick nonseptic (n=22), and healthy foals (n=7) were compared. In addition, results recorded in septic/neutropenic (n=12), septic/non‐neutropenic (n=20), nonseptic/neutropenic (n=8), nonseptic/non‐neutropenic (n=14), and healthy foals (n=7) were also compared. Results: A weak negative correlation was found between MPXI and neutrophil count and between NXM and percentage of band neutrophils. Septic/neutropenic foals had significantly higher MPXI values (median 17.9, minimum–maximum 4.7–42.5) than did septic/non‐neutropenic (1.5, ?24.4 to 22.3), nonseptic/neutropenic (6.6, 0.6–17.9), and nonseptic/non‐neutropenic foals (8.8, ?10.1 to 16.8) but did not differ significantly from controls (12.8, ?8.5 to 20.4). Conclusions: Significant differences in NXM or MPXI were not found when disease groups were compared with controls; however, septic/neutropenic foals had significantly higher median MPXI than other groups of sick foals. Further prospective studies are needed to clarify if this finding is related to decreased neutrophil function or activation in septic/neutropenic foals.     

Reference Value Advisor: a new freeware set of macroinstructions to calculate reference intervals with Microsoft Excel

Abstract: International recommendations for determination of reference intervals have been recently updated, especially for small reference sample groups, and use of the robust method and Box–Cox transformation is now recommended. Unfortunately, these methods are not included in most software programs used for data analysis by clinical laboratories. We have created a set of macroinstructions, named Reference Value Advisor, for use in Microsoft Excel to calculate reference limits applying different methods. For any series of data, Reference Value Advisor calculates reference limits (with 90% confidence intervals [CI]) using a nonparametric method when n≥40 and by parametric and robust methods from native and Box–Cox transformed values; tests normality of distributions using the Anderson–Darling test and outliers using Tukey and Dixon–Reed tests; displays the distribution of values in dot plots and histograms and constructs Q–Q plots for visual inspection of normality; and provides minimal guidelines in the form of comments based on international recommendations. The critical steps in determination of reference intervals are correct selection of as many reference individuals as possible and analysis of specimens in controlled preanalytical and analytical conditions. Computing tools cannot compensate for flaws in selection and size of the reference sample group and handling and analysis of samples. However, if those steps are performed properly, Reference Value Advisor, available as freeware at http://www.biostat.envt.fr/spip/spip.php?article63 , permits rapid assessment and comparison of results calculated using different methods, including currently unavailable methods. This allows for selection of the most appropriate method, especially as the program provides the CI of limits. It should be useful in veterinary clinical pathology when only small reference sample groups are available.     

Canine reference intervals for the Sysmex XT-2000iV hematology analyzer

Background: The laser‐based Sysmex XT‐2000iV hematology analyzer is increasingly used in veterinary clinical pathology laboratories, and instrument‐specific reference intervals for dogs are not available. Objective: The purpose of this study was to establish canine hematologic reference intervals according to International Federation of Clinical Chemistry and Clinical and Laboratory Standards Institute guidelines using the Sysmex XT‐2000iV hematology analyzer. Methods: Blood samples from 132 healthy purebred dogs from France, selected to represent the most prevalent canine breeds in France, were analyzed. Blood smears were scored for platelet (PLT) aggregates. Reference intervals were established using the nonparametric method. PLT and RBC counts obtained by impedance and optical methods were compared. Effects of sex and age on reference intervals were determined. Results: The correlation between impedance (I) and optical (O) measurements of RBC and PLT counts was excellent (Pearson r=.99 and .98, respectively); however, there were significant differences between the 2 methods (Student's paired t‐test, P<.0001). Differences between sexes were not significant except for HCT, PLT‐I, and PLT‐O. WBC, lymphocyte, and neutrophil counts decreased significantly with age (ANOVA, P<.05). Median eosinophil counts were higher in Brittany Spaniels (1.87 × 10⁹/L), Rottweilers (1.41 × 10⁹/L), and German Shepherd dogs (1.38 × 10⁹/L) than in the overall population (0.9 × 10⁹/L). PLT aggregates were responsible for lower PLT counts by the impedance, but not the optical, method. Conclusion: Reference intervals for hematologic analytes and indices were determined under controlled preanalytical and analytical conditions for a well‐characterized population of dogs according to international recommendations.     

Anti-Neospora caninum antibodies in milk in relation to production losses in dairy cattle

A comprehensive field study was carried out with the following objectives: (a) to assess the usefulness of individual and bulk tank milk analysis for determining Neospora caninum serostatus in individual cows and herds, and (b) to study the associations between N. caninum infection status (based on milk testing), and several productive and reproductive parameters in the animals. Antibodies were detected with a commercially available ELISA test (Bio K 192/5). Analysis of paired serum and milk samples from 1134 lactating cows on 38 farms revealed that 97.6% of the ELISA results were coincident, irrespective of whether serum or milk samples were used. Moreover, multiple linear regression analysis revealed that 86.0% of the variations in ELISA values in milk were due to variations in the serum. The measurement of antibodies in bulk tank milk was a good estimator of the herd level status of N. caninum infection, and enabled detection of infection in 94.7% herds with ≥10.0% seropositive cows and/or in all herds with >4% highly seropositive cows. The odds ratio for abortion in seropositive animals was 9.1 times higher than in seronegative animals. The infection serostatus was also a significant risk factor, as the odds ratio for abortion was even higher (12.0 times) in cows categorized as highly seropositive. ELISA values for the bulk milk from 387 randomly selected herds were negatively associated with average milk production. Moreover, milk production losses mainly occurred on farms categorized as highly positive (i.e. herds with ≥20.0% seropositive cows).     

Squamous cell carcinoma of the vagina and cervix in sheep - case report

This report describes the clinical and histopathological characteristics of a squamous cell carcinoma infiltrating the cervix and the vaginal wall, producing reproductive symptoms and subnormal fertility in an adult ewe. Necropsy showed a large (15-cm-long) neoplastic mass infiltrating the vaginal wall and the cervix. Histopathological examination revealed atypical squamous epithelial cords invading the basal membrane and dermis, round anaplastic cells, focal areas of necrosis, keratinisation of isolated cells, and pronounced infiltration by mononuclear cells around the cords. No squamous cell carcinoma of such localisation has been reported from sheep before. In humans, this tumour is the most common gynaecological malignancy in the world.     

Genotyping of potentially zoonotic Giardia duodenalis from exotic and wild animals kept in captivity in Brazil

We have studied the variability of glutamate dehydrogenase (gdh) and small subunit ribosomal (SSU) rRNA coding genes of Giardia species in fecal samples isolated from wild and exotic animals in Brazil, and compared with homologous sequences of isolates from human and domestic animals characterized in previous studies. Cysts of Giardia duodenalis were obtained from feces of naturally infected monkeys (Alouatta fusca) (n=20), chinchillas (Chinchilla lanigera) (n=3), ostriches (Struthio camelus) (n=2) and jaguar (Panthera onca) (n=1). Assemblage AI was assigned to the unique isolate of jaguar. All the samples from monkeys, chinchillas, and ostriches were assigned to Assemblage B. There was little evolutionary divergence between the referred isolates and isolates described elsewhere. The Assemblage B isolates identified in this study were closely related to Assemblage BIV isolated from humans. The molecular identification of Assemblages A and B of G. duodenalis isolates from exotic and wild animals demonstrates that such hosts may be a potential reservoir for zoonotic transmission of G. duodenalis.     

The Pho regulon and the pathogenesis of Escherichia coli

During the course of infection, bacteria must coordinately regulate gene expression in response to environmental stimuli. The phosphate (Pho) regulon is controlled by the two component-regulatory system PhoBR. PhoBR is activated during starvation and regulates genes involved in phosphate homeostasis. Several studies have highlighted the importance of the Pho regulon in bacterial pathogenesis, showing how induction of PhoBR, in addition to regulating genes participating in phosphate metabolism, leads to modulation of many cellular processes. The pleiotropic effects of Pho regulon activation include attenuated virulence and alteration of many virulence traits, including adhesion to host cells and resistance to cationic antimicrobial peptides, acidity and oxidative stresses. This review provides an overview of the relationship between the Pho regulon and virulence in Escherichia coli and illustrates that, in addition to regulating phosphate homeostasis, the Pho regulon plays a key role in regulating stress responses and virulence.     

Growth and development of pepper are affected by humic substances derived from composted sludge

A greenhouse experiment was conducted to evaluate the effects of humic substances extracted from composted sewage sludge on growth, phenological development, and photosynthetic activity of pepper (Capsicum annuum L. cv. Piquillo) plants. Humic substances derived from composted sludge (HSS) were compared with those derived from leonardite (HSL). Two doses of both humic substances were assayed (200 and 500 mg C [L substrate]^–1) and compared with a control (C). HSS showed higher nitrogen content and a higher percentage of aliphatic carbon, as well as a lower content of aromatic and phenolic carbon than HSL. HSS significantly increased plant dry‐matter production (up to 560%), plant height (86%–151%), and leaf area (436%–1397%) during the early stages of pepper development. Net photosynthesis and stomatal conductance increased in the treatments with HSS (up to 48% and 63%, respectively) at the vegetative stage. In addition, HSS accelerated the phenological development of pepper plants, reducing significantly the number of days to flowering and ripening, which occurred 12 and 14 d earlier than in control plants, respectively. In general, the treatments with HSS and HSL did not markedly affect chlorophyll and nutrient concentrations in the leaves. At maturity, only small differences in total fruit yield, number of fruits per plant, and fruit size were observed between amended and control plants. The results suggest that the mechanisms through which HSS affected plant growth and development were not associated with an improved nutrient uptake. Although the identity of the growth‐promoting factors remains to be found, the results suggest that they may be linked to the chemical structure of the humic substances.     

Spatial evaluation of soil salinity using the WET sensor in the irrigated area of the Segura river lowland

The electrical conductivity of the water within the soil pores (EC_p) measured with the WET sensor, appears to be a reliable estimate of soil salinity. A methodology combining the use of the WET sensor along with geostatistics was developed to delimit and evaluate soil salinity within an irrigated area under arid to semiarid Mediterranean climate in SE Spain. A systematic random sampling of 104 points was carried out. The association between EC_p and the saturation‐extract electrical conductivity (EC_se) was assessed by means of correlation analysis. The semivariograms for EC_p were obtained at three different soil depths. Interpolation techniques, such as ordinary kriging and cokriging, were applied to obtain EC_p levels in the unknown places. For each one of the soil depths, a model able to predict EC_se from EC_p was developed by means of ordinary least squares regression analysis. A good correlation (r = 0.818, p < 0.001) between EC_p and EC_se was found. Spherical spatial distribution was the best model to fit to experimental semivariograms of EC_p at 10, 30, and 50 cm soil depths. Nevertheless, cokriging using the EC_p of an adjacent soil depth as an auxiliary variable provided the best results, compared to ordinary kriging. An analytical propagation‐error methodology was found to be useful to ascertain the contribution of the spatial interpolation and ordinary least squares analysis to the uncertainty of the EC_se mapping. This methodology allowed us to identify 98% of the study area as affected by salinity problems within a rooting depth of 50 cm, with the threshold of EC_se value at 2 dS m^–1. However, considering the crops actually grown and 10% potential reduction yield, the soil‐salinity‐affected area decreased to 83%. The use of sensors to measure soil salinity in combination with geostatistics is a cost‐effective way to draw maps of soil salinity at regional scale. This methodology is applicable to other agricultural irrigated areas under risk of salinization.