Identification and localization of the structural proteins of anguillid herpesvirus 1

ABSTRACT: Many of the known fish herpesviruses have important aquaculture species as their natural host, and may cause serious disease and mortality. Anguillid herpesvirus 1 (AngHV-1) causes a hemorrhagic disease in European eel, Anguilla anguilla. Despite their importance, fundamental molecular knowledge on fish herpesviruses is still limited. In this study we describe the identification and localization of the structural proteins of AngHV-1. Purified virions were fractionated into a capsid-tegument and an envelope fraction, and premature capsids were isolated from infected cells. Proteins were extracted by different methods and identified by mass spectrometry. A total of 40 structural proteins were identified, of which 7 could be assigned to the capsid, 11 to the envelope, and 22 to the tegument. The identification and localization of these proteins allowed functional predictions. Our findings include the identification of the putative capsid triplex protein 1, the predominant tegument protein, and the major antigenic envelope proteins. Eighteen of the 40 AngHV-1 structural proteins had sequence homologues in related Cyprinid herpesvirus 3 (CyHV-3). Conservation of fish herpesvirus structural genes seemed to be high for the capsid proteins, limited for the tegument proteins, and low for the envelope proteins. The identification and localization of the structural proteins of AngHV-1 in this study adds to the fundamental knowledge of members of the Alloherpesviridae family, especially of the Cyprinivirus genus.     

Bovine neonatal pancytopenia in German Holstein calves

Profiles of blood cell counts were evaluated for 15 calves from three different farms. These calves showed petechia in the mucous membranes and in the skin and prolonged secondary bleeding after puncture. The clinical course of the disease could be observed in eleven calves. With exception of one case, the blood cell counts indicated a severe anaemia, leukocytopenia and thrombocytopenia. Out of these 15 calves, six calves survived and the other nine calves died or had to be euthanized due to the severity of the disease. Necropsy of these nine calves revealed petechia in the skin, subcutis, muscles, in inner organs and all serous membranes. Pathohistological examination showed a depletion of the bone marrow and lymphatic tissue in eight calves. These findings confirmed the diagnosis of bovine neonatal pancytopenia (BNP) for eight of these nine calves. Bluetongue virus serotype 8 was tested negatively using PCR. Bovine virus diarrhoea virus (BVDV) was negatively tested using immunofluorescence and cell culture and salmonella species were negatively tested in seven dissected calves. A cluster of toxins was negatively tested in one of the dissected calves. All 15 calves had high antibody titres for BVDV. The BVDV-antibody titres from twelve dams with affected calves were positive in six cases and not detectable in the other six cases. In three of the six dams with not detectable BVDV-antibody titres, calves were fed with colostrum of a further dam with high BVDV-antibody titres. In the further three dams without detectable BVDV-antibody titres, we could not ascertain which colostrum has been fed to the calves. BVDV-specific antigen could not be detected in any of the samples from the calves and dams tested. Using the activity of the gamma-glutamyl-transferase, we assumed a sufficient supply with colostrum for the examined calves.The cause for the occurrence of these BNP cases was due to bone marrow depletion.The reason for the bone marrow depletion remained unclear. However, it was obvious that the BNP described here is highly likely caused by colostrum from cows with positive BVDV-antibody titres.     

First morphological characterization of 'Candidatus Mycoplasma turicensis' using electron microscopy

At least three haemotropic mycoplasmas have been recognized in cats: Mycoplasma haemofelis (Mhf), 'Candidatus Mycoplasma haemominutum' (CMhm) and 'Candidatus M. turicensis' (CMt). The latter was originally identified in a Swiss pet cat with haemolytic anaemia and shown to be prevalent in domestic cats and wild felids worldwide using molecular methods. So far, there has been no confirmatory morphological evidence of the existence of CMt presumably due to low blood loads during infection while CMhm has only been characterized by light microscopy with discrepant results. This study aimed to provide for the first time electron microscopic characteristics of CMt and CMhm and to compare them to Mhf. Blood samples from cats experimentally infected with CMt, CMhm and Mhf were used to determine copy numbers in blood by real-time PCR and for transmission and scanning electron microscopy. High resolution scanning electron microscopy revealed CMt and CMhm to be discoid-shaped organisms of 0.3 μm in diameter attached to red blood cells (RBCs). In transmission electron microscopy of CMt, an oval organism of about 0.25 μm with several intracellular electron dense structures was identified close to the surface of a RBC. CMhm and CMt exhibited similar morphology to Mhf but had a smaller diameter. This is the first study to provide morphological evidence of CMt thereby confirming its status as a distinct haemoplasma species, and to present electron microscopic features of CMhm.     

Influence of the Expert Effect on Cultural Models

We examined hunters’ perceptions of fauna to see if expert hunters and other hunters perceive wildlife abundance similarly. We used cultural consensus analysis (CCA) to assess the knowledge of 25 hunters in the Bolivian Amazon about the abundance of 38 animals. CCA indicated highly shared beliefs among hunters concerning wildlife abundance (average agreement = .62). However, expert hunters (as judged by their reported successful hunts of rare species, having hunted recently, and consuming more game in their diet) perceived more animals as abundant than did non-experts, although they all shared the same model. Since the expert hunters did not always agree on which species was more abundant, they had low cultural knowledge scores in CCA results. These experts may be unwilling to curtail hunting efforts on key species that they perceive to be abundant.     

Characterization of a feline homologue of the alphaE integrin subunit (CD103) reveals high specificity for intra-epithelial lymphocytes

The characteristics of a feline homologue of the alphaE integrin (CD103), defined by two murine monoclonal antibodies, Fe7.1B8 (IgG1) and Fe7.2D8 (IgG1), are described. These antibodies recognized 75% of intra-epithelial (range 59-88%) and 40% of lamina proprial (range 28-46%) T cells of the intestinal mucosal tissue of the small intestine in contrast with approximately 2% of peripheral blood lymphocytes. Both antibodies immunoprecipitated a 180 kDa protein from biotinylated feline intra-epithelial mucosal leukocytes consistent with the alphaE integrin subunit in conjunction with a 120 kDa protein consistent with the beta7 subunit. The nucleotide sequence of feline alphaE integrin, generated from molecular cloning of the feline alphaE encoding cDNA, is also reported. This feline molecule shares 72% sequence homology with human and 69% homology with murine and rat counterparts. Homology includes the presence of an X (extra) domain, that appears unique to alphaE molecules as described for human, rat and mouse, as well as areas of homology common to other alpha integrins. Of note is a typical I (inserted) domain, the presence of seven repeat regions, and highly conserved sequences in the cytoplasmic tail. Transfection studies demonstrated that both antibodies recognized an extracellular component which encompassed the X and I domains of the cloned alphaE integrin subunit. These studies demonstrate that the pattern of tissue distribution, biochemical characteristics, and cDNA sequence of the feline alphaE integrin subunit are largely similar to that described for other species.     

Prevalence of systolic hypertension in cats with chronic renal failure at initial evaluation

OBJECTIVE: To determine prevalence of systolic hypertension and associated risk factors in cats with chronic renal failure evaluated in first-opinion practice. DESIGN: Prospective study. ANIMALS: 103 cats with chronic renal failure. PROCEDURE: Systolic arterial blood pressure (SABP) was measured with a noninvasive Doppler technique, and cats that had SABP > 175 mm Hg on 2 occasions or that had SABP > 175 mm Hg and compatible ocular lesions were classified as hypertensive. Information from the history (previous treatment for hyperthyroidism, age), physical examination (sex, body weight), routine plasma biochemical analyses (creatinine, cholesterol, potassium, sodium, chloride, and calcium concentrations), and thyroid status were evaluated as potential risk factors for systolic hypertension. Variables associated with systolic hypertension were evaluated by use of logistic regression. RESULTS: 20 (19.4%; 95% confidence interval, 13 to 28%) cats had systolic hypertension. Plasma potassium concentration was significantly and inversely associated with systolic hypertension. CONCLUSIONS AND CLINICAL RELEVANCE: Prevalence of systolic hypertension, although clinically important, was lower than that reported previously. The cause of the inverse association between systolic hypertension and plasma potassium concentration is not yet known.     

Effects of Bacillus cereus var. toyoi on immune parameters of pregnant sows

Changing immune parameters during pregnancy have previously been reported in humans and cattle, and have been suggested to contribute to increased susceptibility to infections. However, data regarding immune parameters during pregnancy in sows are rare. In this study, we investigated the peripartal immune status of sows using phenotypical (FACS analysis) as well as functional (proliferation assays, cytokine analysis) parameters of peripheral blood mononuclear cells (PBMCs) in pregnant sows. In previous studies, we reported a modulation of the immune system after feed supplementation of the probiotic Bacillus cereus var. toyoi in piglets [Schierack, P., Wieler, L.H., Taras, D., Herwig, V., Tachu, B., Hlinak, A., Schmidt, M.F., Scharek, L., 2007. Bacillus cereus var. toyoi enhanced systemic immune response in piglets. Vet. Immunol. Immunopathol. 118, 1–11]. Here, we extended these previous studies to include investigations of possible probiotic effects on the peripartal immune status of sows and their reproductivity. We show that immune parameters of sows change during pregnancy, the proliferative response of PBMCs to several bacterial antigens in control animals decreased from days 90 to 30 ante partum. Relative numbers (%) of CD3+CD8+, CD4+, cytotoxic T, CD14+ and CD21+ cells were reduced compared to non-pregnant sows. In contrast, the proliferative response of PBMCs of probiotic-treated sows increased during pregnancy. Bacterial antigens primarily stimulated the proliferation of naïve CD21+ cells and the relative CD21+ cell numbers were elevated in the probiotic group in the absence of effects on other immune cell populations. The clinical and microbial status of both control and probiotic sows was similar, excluding pre-existing health problems or infections as responsible for the immunological changes, and feed supplementation also had no significant effects on reproductivity. The results suggest that the probiotic B. cereus var. toyoi can alter the proliferative response of lymphocytes and affects the immune cell population ratios of pregnant sows. How and to what extent this may affect health and reproductivity should be the focus of further studies.     

The effect of selected microbial strains on internal milieu of broiler chickens after peroral administration

The aim of this work was to investigate the effect of probiotic preparation with multi-strains composition (Lactobacillus delbrueckii ssp. bulgaricus LAT 187, Lactobacillus acidophilus LAT 180, Lactobacillus helveticus LAT 179, Lactobacillus delbrueckii ssp. lactis LAT 182, Streptococcus thermophilus LAT 205 and Enterococcus faecium E 253) on internal milieu, antioxidant status and body weight of broiler chickens. Chickens were divided into three groups (C, control; E1 and E2, experimental groups). Experimental chickens received the probiotic preparation in drinking water. Probiotic preparation caused a significant increase of serum calcium and potassium content. In the group with higher dose of probiotic strains serum triglycerides level decreased. Total antioxidant status in groups with addition of probiotic strains showed higher values in comparison to control group. Serum albumin level was found to increase after consumption of probiotic preparation in group with higher dose of probiotic strains. Probiotic strains improved body weight in last observed weeks of feeding.     

Letter to the Editor

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