Brucella intracellular life: from invasion to intracellular replication

Brucella organisms are pathogens that ultimate goal is to propagate in their preferred niche, the cell. Upon cell contact the bacteria is internalized via receptor molecules by activating small GTPases of the Rho subfamily and by a moderate recruitment of actin filaments. Once inside cells, Brucella localizes in early phagosomes, where it avoids fusion with late endosomes and lysosomes. These early events require the control of Rab small GTPases, and cytokines such as the G-CSF. Then, the bacterium redirects its trafficking to autophagosomes and finally reaches the endoplasmic reticulum, where it extensively replicates. Some of the bacterial molecular determinants involved in the internalization and early events after ingestion are controlled by the BvrS/BvrR two component regulatory system, whereas the intracellular trafficking beyond this early compartments are controlled by the VirB type IV secretion system. Once inside the endoplasmic reticulum, Brucella extensively replicates without restricting basic cellular functions or inducing obvious damage to cells. The integrity of Brucella LPS on the bacterial surface is one of the required factors for Brucella intracellular survival, and therefore for virulence.     

In vitro isolation and characterization of bovine Neospora caninum in Korea

The brains of nine aborted bovine fetuses and two newborn calves born from dams suspected to be infected with Neospora caninum were homogenized and inoculated into Vero cells. All fetuses and calves were from cows determined as seropositive to N. caninum by an IFA test. Sera and thoracic fluids of all fetuses and calves also revealed high antibody titer to N. caninum by IFAT ranging from 1:800 to 1:3200. N. caninum was isolated from the brains of one aborted fetus and one newborn calf when the brain homogenates were grown continuously in Vero cell culture. N. caninum tachyzoites, giemsa-positive, were first observed on Days 45 and 56 postinoculation in the newborn calf and the aborted fetus, respectively. The isolates (KBA-1 and KBA-2) were morphologically and ultrastructurally similar to previously published Neospora isolates. The isolated parasites were confirmed as N. caninum by means of the antigenic reactivities, immunostaining, PCR and southern blotting, and electron microscopy.     

Development and validation of a radioimmunoassay for thyrotropin in cattle.

In mammals, thyrotropin, or thyroid-stimulating hormone (TSH), assay is used for the diagnosis of primary hypothyroidism. Hypothyroidism is the most common type of thyroid disorder in cattle. The aim of this study was to develop and validate, under physiologic and pathologic conditions, a radioimmunoassay (RIA) for bovine TSH (bTSH). Double RIA was performed with purified bTSH and specific bovine antiserum. Laboratory validation included research of minimal detection limit, accuracy, and reproducibility. The physiologic validation included a thyrotropin-releasing hormone (TRH) challenge performed on euthyroid cows and a follow-up of bTSH concentration over a 24-hour period. Furthermore, bTSH concentration was assayed in a large population of healthy dairy and beef cows to define reference interval. The pathologic validation was made by assaying bTSH and thyroid hormones on healthy and goitrous newborn calves. The minimum detection limit (MDL) for bTSH assay was 1.3 microU/ml. The recovery was 101% to 106%. The intra- and interassay coefficients of variation (CVs) ranged from 5% to 11% and 11% to 15%, respectively. The RIA covered the whole range of physiologic bTSH values, as shown by bTSH values induced by TRH-challenge. A pulsatile secretion of bTSH was observed, accompanied by a diurnal variation with lower night values than day values. Reference intervals of bTSH ranged from 1.3 to 13.0 microU/ml for beef and dairy breeds. Finally, bTSH easily discriminated goitrous newborn calves from healthy ones, leading to the definition of a cutoff value of 35 microU/ml. The bTSH assay positively reacted to physiologic and pathologic conditions. The accuracy and precision of the RIA were satisfying.     

C-reactive protein, tumor necrosis factor α, and interleukin-6 in dogs with pyometra and SIRS

Objective: To determine the frequency of the systemic inflammatory response syndrome (SIRS) in canine pyometra and to evaluate the relationship between C‐reactive protein (CRP), tumor necrosis factor α (TNFα), interleukin‐6 (IL‐6), and SIRS. Design: Prospective clinical study. Setting: Veterinary teaching hospital. Animals: Fifty‐three clinical cases of canine pyometra and 19 healthy control bitches. Interventions: Upon admission to the veterinary hospital, history and physical examination findings, including previously defined clinical SIRS parameters, were documented. Blood samples were obtained for hematology and biochemical tests and for CRP, TNFα, and IL‐6 analysis. The diagnosis of pyometra was confirmed by histopathology of the uterus after ovariohysterectomy. After surgery, clinical SIRS parameters, length of hospitalization, and mortality were recorded. Measurements and main results: Pyometra dogs were grouped as SIRS positive (30/53; 57%) or SIRS negative (23/53; 43%). Logistic regression showed that CRP was the only parameter that significantly related to SIRS apart from the clinical criteria that define this syndrome. The mortality rate was low (2/53; 3.8%), and conclusions regarding association with SIRS could not be drawn. A positive SIRS status, high plasma CRP concentration, and high body temperature were variables that related to increased morbidity reflected by the length of hospitalization. Conclusions: SIRS was seen in 57% of canine pyometra cases and a positive SIRS status showed a positive association with prolonged hospitalization. The mortality rate was low (3.3%) among SIRS positive dogs, indicating that progression to multiple organ dysfunction syndrome (MODS) rarely occurs in surgically treated cases of pyometra. CRP was associated with SIRS and with prolonged hospitalization. Further studies of plasma CRP may be warranted in canine intensive care cases susceptible to development of SIRS and MODS.     

Pasteurella multocida isolation in a horse with retropharyngeal infection

Retropharyngeal infections in horses normally induce local painful swelling of the retropharyngeal area, which may lead to dyspnea, dysphagia, and systemic manifestations. Differential diagnosis of local painful swelling of the retropharyngeal area includes retropharyngeal lymph node infection, neoplasm, cellulitis, hematoma, guttural pouch empyema, parotiditis, and jugular thrombosis. Apart from Streptococcus equi ssp. equi, other bacteria are rarely reported as a cause of retropharyngeal abscesses. The reason for this might be a lack of specific sampling to identify the causative agent. This work deals with a case of retropharyngeal infection in an 11-year-old Standardbred stallion with acute depression, fever, tachycardia, asymmetric painful swelling in the throat area, ptyalism, and respiratory distress. Endoscopy, radiography, ultrasonography, blood analysis, and cytological examination of a puncture sample taken from the throat mass were consistent with a pyogenic to pyogranulomatous retropharyngeal inflammation. The clinical evolution was initially satisfactory in response to treatment with nonsteroidal anti-inflammatory drugs and antibiotics, but clinical signs relapsed twice, each time a few weeks after cessation of antibiotic therapy. The bacteriologic finding in this case was unusual and consisted of the isolation of a Pasteurella multocida strain that was obtained after the second relapse (ie, 79 days after initial admission), using a brain heart infusion (BHI) medium, and after two successive negative bacteriological cultures performed on day one of clinical signs and at the first relapse of clinical signs, respectively.     

Effect of cultural practices on the development of arabica coffee berry disease,caused by <Emphasis Type="Italic">Colletotrichum kahawae</Emphasis>

In the high altitude regions of Africa, coffee berry disease (CBD), caused by Colletotrichum kahawae, is the main constraint for arabica coffee (Coffea arabica) production. However, certain agricultural practices can reduce losses caused by the disease and thereby promote optimum production. On small family farms in Cameroon, mixed cropping with fruit trees, intercropping with food crops and maintenance pruning of coffee trees are very widespread agricultural practices that can affect CBD epidemics. Consequently, an epidemiological study was conducted to assess how cultural practices affected the disease in an arabica coffee smallholding in Cameroon. The disease was monitored on a weekly basis over four successive years (2002–2005) on coffee trees in diverse cultural situations. Cultural practices likely to reduce losses due to CBD were identified. The infection rate was significantly lower on coffee trees grown intensively than on coffee trees grown in the traditional manner. Coffee trees located under the shade of fruit trees were significantly less attacked than those located in full sunlight. In addition, berries on the leafless parts of branches, near the main trunk of the coffee tree, were less infected than those on leafy sections. These results show that maintenance pruning, removal of mummified berries, and mixed cropping with shade plants are cultural practices which create environmental conditions that limit CBD development.     

A variance-covariance structure to take into account repeated measurements and heteroscedasticity in growth modeling

This study proposes a within-subject variance-covariance (VC) structure to take into account repeated measurements and heteroscedasticity in a context of growth modeling. The VC structure integrates a variance function and a continuous autoregressive covariance structure. It was tested on a nonlinear growth model parameterized with data from permanent sample plots. Using a stand-level approach, basal area growth was independently modeled for red spruce (Picea rubens Sarg.) and balsam fir [Abies balsamea (L.) Mill.] in mixed stands. For both species, the implementation of the VC structure significantly improved the maximum likelihood of the model. In both cases, it efficiently accounted for heteroscedasticity and autocorrelation, since the normalized residuals no longer exhibited departures from the assumptions of independent error terms with homogeneous variances. Moreover, compared with traditional nonlinear least squares (NLS) models, models parameterized with this VC structure may generate more accurate predictions when prior information is available. This case study demonstrates that the implementation of a VC structure may provide parameter estimates that are consistent with asymptotically unbiased variances in a context of nonlinear growth modeling using a stand-level approach. Since the variances are no longer biased, the hypothesis tests performed on the estimates are valid when the number of observations is large.     

The dehesa: an agrosilvopastoral system of the Mediterranean region with special reference to the Sierra Morena area of Spain

A multidisciplinary team undertook a six-year investigation on the agro-ecological and socio-economic aspects of the dehesa system in the Sierra Norte area, a part of the Sierra Morena of the Sevilla Province, Western Andalusia, Spain. This paper summarizes its findings on the various agro-ecological features of the system and evaluates the system's functional aspects vis-a-vis the trends and developments in land use in the region over the past few centuries. The synergistic effects of tree cover on understorey grassland vegetation is discussed in terms of soil fertility build-up and favourable micro-climatic and hydrological features, and the potential use of this information in future research and development programmes to improve the dehesa system in the Mediterranean region is outlined.     

Impact of successive drought and re-watering cycles on growth and specific leaf area of two Populus x canadensis (Moench) clones, 'Dorskamp' and 'Luisa_Avanzo'

Responses to successive drought and re-watering cycles (1-3 cycles) were compared in greenhouse-grown cuttings of Populus x canadensis (Moench) clones, 'Luisa_ Avanzo' and 'Dorskamp.' Total leaf number increment rate, duration of leaf expansion, total and individual leaf area expansion rates and stomatal conductance were recorded periodically during the experiment. Soil water content (SWC) and predawn leaf water potential (Psi(WP)) were measured four times during each drought cycle. In parallel, relative leaf water content (RWC) and specific leaf area (SLA) were estimated on leaves collected from the top to bottom of each cutting. Under well-watered conditions, 'Luisa_Avanzo' and 'Dorskamp' differed in their patterns of leaf area expansion. Although duration of leaf expansion was similar between clones, 'Luisa_ Avanzo' exhibited higher total leaf number increment rates and individual leaf area increases than 'Dorskamp.' As a result, 'Luisa_Avanzo' cuttings reached larger individual and total leaf areas than 'Dorskamp.' 'Dorskamp' leaves had lower SLA than 'Luisa_Avanzo' leaves. In response to successive drought cycles, both clones underwent decreases in total leaf number increment rates and in total leaf area expansion rates, but both whole-plant and individual leaf areas were drastically reduced only in 'Luisa_Avanzo.' 'Dorskamp' maintained a constant leaf area as a result of an increase in the duration of leaf expansion during drought and a significant stimulation of individual leaf area expansion rate and total leaf number increment rate in response to re-watering. Drought caused a greater decrease in SLA in 'Luisa_Avanzo' than in 'Dorskamp.' Expanded leaves of 'Dorskamp' were constitutively dense or thick, or both, whereas leaves of 'Luisa_Avanzo' became dense or thick, or both, only in response to drought. In both clones, re-watering caused partial recovery of SLA to control values. Our data confirm previous field and greenhouse observations that 'Dorskamp' is more tolerant to successive drought and re-watering cycles than 'Luisa_Avanzo.' We conclude that this difference is mainly attributable to the plasticity of 'Dorskamp' after re-watering, rather than to clonal differences in drought responses.