Early acute depletion of lymphocytes in calicivirus-infected adult rabbits

Rabbit Haemorrhagic Disease (RHD) is a lethal infection caused by calicivirus that kills 90% of the infected adult rabbits within 3 days. The calicivirus replicates in the liver and causes a fulminant hepatitis. Most studies on the pathology of RHD have been focused on the fulminant liver disease. This may not be the only mechanism in the pathogenesis of RHD: calicivirus infection may also induce leukopenia in the infected adult rabbits. We show now by flow cytometry analysis that the calicivirus induces an early decrease in B and T cells, in both spleen and liver. The depletion of B and T cells was associated with apoptosis labelled by annexin V. These changes occurred in rabbits before they showed enzymatic evidence of liver damage and persisted after liver transaminase values were very high. We conclude that depletion of lymphocytes caused by the calicivirus infection precedes or attends liver damage. The relative contribution of this lymphocyte depletion for the pathogenesis of the fatal calicivirus infection of rabbits remains to be investigated.     

Low genetic variability in Sclerotinia sclerotiorum populations from common bean fields in Minas Gerais State,Brazil, at regional,local and micro‐scales

Sclerotinia sclerotiorum, causal agent of white mould, is the most destructive and widely distributed soilborne pathogen of common bean during the autumn–winter season in Brazil. Nevertheless, little is known about the genetic structure of the pathogen population. Microsatellite (SSR) markers and mycelial compatibility groups (MCGs) were used to characterize 118 isolates collected from 20 bean fields located in the most important growing regions of Minas Gerais State (MG). Additionally, the genetic variability among 10 isolates obtained from a single sclerotium was investigated in 10 different sclerotia. Seventy SSR haplotypes and 14 MCGs were identified among the 118 isolates. The genetic differences within bean growing areas accounted for most of the genetic variation (72%). Despite the relatively high genotypic diversity, the SSR loci were at linkage disequilibrium. Moreover, 70% of the isolates were assigned to only two MCGs, and haplotypes of a given MCG were closely related. The discriminant analysis of principal components revealed five groups. There was strong genetic differentiation between isolates collected in one municipality in southern MG when compared to other regions. Common bean resistance to white mould should be assessed with representative isolates of the five genetic groups and, if possible, of the different MCGs detected in the present study. One to five haplotypes were detected among the 10 isolates obtained from a single sclerotium. Therefore, in order to ensure genetic identity of an isolate, hyphal tip or monoascosporic isolates should be used.     

Comparison of postoperative pain after ovariohysterectomy by harmonic scalpel-assisted laparoscopy compared with median celiotomy and ligation in dogs

OBJECTIVE: To compare the effects of postoperative pain after ovariohysterectomy by harmonic scalpel-assisted laparoscopy (HALO) and traditional ovariohysterectomy (OVH) in dogs. STUDY DESIGN: A randomized, blinded, prospective study. SAMPLE POPULATION: Sixteen, purpose-bred, intact female, Beagle dogs. METHODS: Dogs were divided into 2 groups: Group 1 (8 dogs), which had OVH by HALO, and Group 2 (8 dogs), which had traditional OVH. Physiologic data, abdominal nociceptive threshold scores, and University of Melbourne pain scores (UMPS) were recorded at 2, 6, 12, 24, 48, and 72 hours after surgery. Blood samples for measurement of plasma cortisol, glucose, and creatine phosphokinase (CPK) concentrations were collected at the time of the incision, and 2, 6, 12, 24, 48, and 72 hours after surgery. RESULTS: No significant surgical complications occurred. The HALO mean surgical time was significantly longer (55.7 minutes) than traditional OVH (31.7 minutes). No significant differences were observed between groups for the pain measures of heart rate, respiratory rate, temperature, CPK, and glucose concentrations. The OVH group had significantly higher mean plasma cortisol levels at hour 2 after surgery than the HALO group (P=.0001). The mean UMPS were significantly higher in OVH than the HALO group at all postoperative times (P=.0001). The mean nociceptive threshold measurements revealed significantly higher tolerated palpation pressures in HALO than OVH at all postoperative times, except hour 72 (P=.0002). CONCLUSIONS: Dogs appeared to be in less pain with HALO than OVH. The harmonic scalpel coagulated ovarian and uterine vessels completely with minimal collateral damage to surrounding tissues. CLINICAL RELEVANCE: HALO is a safe alternative to OVH and offers a minimally invasive and less painful method of surgery.     

Risk factors associated with feline urolithiasis

Urinary tract diseases are among the main reasons for consultation in veterinary clinics and hospitals. It affects animals of any age, breed and gender. Among the diseases that affect this system, urolithiasis is the second largest cause of clinical signs compatible with feline urinary tract disease. The term urolithiasis refers to the presence of uroliths in any region of the urinary tract, but it is more commonly seen in the bladder and urethra. Uroliths are classified based on the type of mineral present in their composition, therefore, quantitative and qualitative analyzes are important for a better therapeutic approach. The animals may suffer from the disease and be asymptomatic, or show nonspecific clinical signs, making the diagnosis difficult. The disease should not be seen as a single problem, but as a consequence of various disorders. As dietary, metabolic, genetic and infectious causes, as well as factors that potentiate the chance of development of uroliths such as breed, age, sex, age range, obesity, sedentary lifestyle, geographic region and climate. Thus, the knowledge of the factors that influence the formation of uroliths, as well as the understanding of the pathophysiology, are key elements for better alternatives of therapy and prevention. The recognition of these factors helps to identify susceptible populations, minimizing exposure and increasing the protection factors, which facilitates the diagnosis and treatment of patients with urolithiasis. The objective of this paper is to present the main risk factors involved in the formation of urinary lithiasis in felines.     

Cannibalism among <Emphasis Type="Italic">Lithodes santolla</Emphasis> (Molina 1782) juveniles: effect of stocking density,stage and molt condition

Losses of king crabs (deaths) in massive rearing conditions are frequently attributed to cannibalism. As several factors could influence cannibalism intensity during culture, the aim of the present study was to determine the effect of molt condition, stage and density on cannibalism among Lithodes santolla juveniles. Comparisons between intermolt and molt conditions and between C1 and C2 stages during intermolt were performed at three densities (6, 12 and 18 individuals per flask; that is equivalent to 236, 472 and 708 individuals m^?2). After 16 days of juvenile coexistence, cannibalism (injured + dead) was higher during molting (33.9 ± 4.5 %) than intermolt (21.7 ± 3.9 %), even though only 36 % of crabs had molted. Although cannibalism was similar in C1 and C2 stages, the proportion of dead animals was higher by a factor of three in C1 stage, probably due to the fact that animals reach the first juvenile stage with low energetic reserves, after the lecithotrophic larval development. Density was the main factor influencing cannibalism in our experiments. Higher densities promoted higher deaths, while agonistic interactions evidenced by injured crabs remained constant. Walking appendages were lost more often than chelipeds during first agonistic encounters. We recommend the use of intermediate densities (450 crabs m^?2 at most), the early detection of exuviae and the sorting of crabs after molting, for L. santolla rearing.     

Development of a potentiometric method to measure the resistance to oxidation of white wines and the antioxidant power of their constituents

This work describes a new potentiometric method to evaluate the resistance to oxidation of white wines. Reduction and oxidation titrations were made, and coefficient of variation obtained were 10.87 and 2.65%, respectively. The antioxidant powers of ascorbic acid (Aas) and sulfur dioxide (SO(2)) were evaluated by this method, SO(2) proving to be much less active in this respect than ascorbic acid. The two agents did not demonstrate any antioxidant synergy. A relationship between oxygen present and ascorbic acid was found by the proposed method (1 mmol of O(2) <--> 0.84 mmol of Aas). This method enables the distinction of different wines on the basis of their resistance to oxidation.     

The use of sewage sludge as soil amendment. The need for an ecotoxicological evaluation

Background, aim, and scope  Sewage sludge use in agriculture should be limited by the presence of metals and other persistent environmental pollutants. The present study aims to contribute for the definition of a test battery of ecotoxicological assays that allows a proper ecotoxicological characterization of sludges, providing information on their potential hazard and identified “safe” application levels. Materials and methods  Three sludges from distinct sources (urban, olive-processing, and electroplating industries) were tested using avoidance and reproduction tests with earthworms (Eisenia andrei) and springtails (Folsomia candida) and plant growth tests with turnips (Brassica rapa) and oats (Avena sativa). Different soil–sludge mixture concentrations mimicking recommended/realistic field dosages were tested. Results  Only the sludge from the electroplating industry induced an avoidance response from the earthworms (EC₅₀ = 0.4 t/ha) and collembolans (no observed effect concentration (NOEC) = 15 t/ha). This sludge was the only sludge responsible for any effect on the reproductive output of the earthworms (EC₅₀ = 7.74 t/ha). Regarding collembolans, none of the sludges tested caused any significant decrease in reproduction. In higher plant tests, the two industrial sludges were toxic, causing a decrease growth in both species. The EC₂₀ values determined for B. rapa were 20.3 and 24.2 t/ha and for A. sativa 14.7 and 16.2 t/ha for sludges from olive-processing and electroplating industries, respectively. Discussion  The metal loadings of the different test sludges could partially explain the results obtained. The toxicity of the test sludge from electroplating industry observed on the tested invertebrates and plants could be explained by the high amount of total chromium from which 22.3% was in the most toxic oxidation state—Cr(VI). However, the toxicity caused by the sludge from the olive-processing industry in the test plants could be attributed to the presence of other compounds (not measured in this study) since the metal content was not high enough to induce such an effect. The absence of toxicity showed by the urban test sludge was in agreement with its low levels of metals. Conclusions  The response of the different test organisms and end points varied according to the sludge type. The urban sludge was non-toxic whereas the sludge from the electroplating industry caused a toxic effect on almost all parameters measured (avoidance behavior of both test organisms, reproduction of earthworms, and growth of both plant species). Sludge from the olive-processing industry only caused a toxic effect on growth of both plant species. By analyzing the sensitivity of the different parameters for the most toxic sludge, it was found that avoidance and reproduction were more sensitive than plant growth, whereas plant seed germination was not sensitive at all. Recommendations and perspectives  The ecotoxicological evaluation of wastes can be used as an environmental safety control of sludge use in agriculture. A tiered approach could be adopted for this purpose, incorporating avoidance tests in the first tier (screening level) and reproduction and plant growth tests in a second tier. But more evidence aiming to define the most suitable ecotoxicological test battery for specific sludges with a different contamination profile is still needed.     

Effect of the US3 protein of bovine herpesvirus 5 on the actin cytoskeleton and apoptosis

The US3 protein is a unique protein kinase only present in the Alphaherpesvirinae subfamily of the herpesviruses. Studies performed with several alphaherpesviruses demonstrated that the US3 protein is involved in cytoskeleton modifications during viral infection and displays anti-apoptotic activity. However, the US3 protein of BoHV-5 has not been studied up to now. As reported for other alphaherpesviruses, our results showed that BoHV-5 US3 confers resistance against apoptosis and induces cytoskeletal reorganization leading to cell rounding, actin stress fiber breakdown and cell projections that interconnect cells. The expression of a kinase-dead version of BoHV-5 US3 showed that the anti-apoptotic activity and the induction of cell projections are kinase-dependent whereas kinase activity is not absolutely required for actin stress fiber breakdown. Besides, the kinase-dead version of US3, but not the wild type protein, was found excluded from the nucleus. These results constitute the first report on the BoHV-5 US3 functions, and highlight that there are functional differences and similarities among US3 proteins of different alphaherpesviruses.