Real-time PCR investigation of feline leukemia virus proviral and viral RNA loads in leukocyte subsets

Cats exposed to feline leukemia virus (FeLV), a naturally occurring gammaretrovirus develop either progressive or regressive infection. Recent studies using analyses with enhanced sensitivity have correlated loads throughout FeLV with the clinical outcome, though remarkably, during the acute phase of infection, proviral and viral RNA burdens in the peripheral blood do not differ between groups. We hypothesized that viral loads in specific leukocyte subsets influence the infection outcome. Using a method established to determine the proviral and cell-associated viral RNA loads in specific leukocyte subsets, we evaluated viral loads in eleven FeLV-exposed specific pathogen-free (SPF) cats 2.5 years post-infection. Six cats had undergone regressive infection whereas five were persistently viremic. Aviremic cats had lower total proviral blood loads than the persistently infected cats and FeLV proviral DNA was shown to be integrated into genomic DNA in four out of four animals. Lymphocytes were predominantly infected vs. moncytes and granulocytes in aviremic cats. In contrast, persistently viremic cats were provirus-positive in all leukocyte subsets. The acute phase kinetics of FeLV infection were analyzed in two additional cats; an early lymphoreticular phase with productive infection in lymphocytes in both cats and in monocytes in one cat was followed by infection of the granulocytes; both cats became persistently infected. These results indicate that FeLV persistent viremia is associated with secondary viremia of bone marrow origin, whereas regressive cats only sustain a non-productive infection in low numbers of lymphocytes.     

Molecular markers for Ve1 and Ve2 Verticillium resistance genes from Italian tomato germplasm

The so‐called Rosa (= pink) tomatoes, which are typically grown in the Southern Italian area, are characterized by the pink colour of the fruit, due to the gene y, colourless fruit skin. In a preliminary survey, it was found that among these Rosa tomatoes there were some ‘Rosa di Sorrento’ local landraces showing resistance to Verticillium wilt (race 1). In tomato, resistance to race 1 of V. dahliae and V. albo‐atrum is conferred by two strictly associated genes, Ve1 and Ve2, which independently confer resistance to the same pathogen. The development of two new markers for Ve1 and Ve2, based respectively on selective allele‐specific PCR amplification and on a PCR amplification followed by enzymatic restriction, is reported. These two markers allow the identification of both allelic forms at the Ve loci and they are of potential interest for use in marker‐assisted selection. Furthermore, ‘Rosa di Sorrento’‐resistant lines have the same resistance alleles as those found in the Ve‐resistant cultivars.     

Corneal invasion by hemangiosarcoma in a horse

A 15-year-old gray Arabian gelding presented for evaluation of a lateral limbal mass extending across approximately 30% of the cornea. Grossly, the raised mass appeared nonpigmented, smooth, and irregular in shape, with an area of central necrosis and serosanguinous discharge. The mass was removed via lamellar keratectomy and histopathologic evaluation revealed features characteristic of hemangiosarcoma (HSA), including irregular vascular channels lined by a plump spindle cell population. Immunohistochemistry results showed that the neoplastic cells lining the vascular channels present diffuse and strong cytoplasmic reaction with von Willebrand Factor and the perivascular spindle cells exhibit moderate cytoplasmic reaction for smooth muscle actin. A lack of cytokeratin staining definitively excluded a diagnosis of atypical squamous cell carcinoma. Smooth muscle actin staining of the perivascular cells adjacent to the neoplastic endothelial cells is not a feature commonly described in HSA and has not been reported in previous cases of equine HSA. The horse remained in good health 21 months postkeratectomy and has exceeded the survival time of previously documented equine ocular HSA cases where more extreme surgical excision was performed.     

Exposure of cats to low doses of FeLV: seroconversion as the sole parameter of infection

In felids, feline leukemia virus (FeLV) infection results in a variety of outcomes that range from abortive (virus readily eliminated and never detectable) to progressive infection (persistent viremia and viral shedding). Recently, a novel outcome was postulated for low FeLV infectious doses. Naïve cats exposed to faeces of persistently infected cats seroconverted, indicating infection, but remained negative for provirus and p27 antigen in blood. FeLV provirus was found in some tissues but not in the bone marrow, infection of which is usually considered a necessary stage for disease progression. To investigate the impact of low FeLV doses on young cats and to test the hypothesis that low dose exposure may lead to an unknown pathogenesis of infection without involvement of the bone marrow, 21 cats were infected oronasally with variable viral doses. Blood p27, proviral and viral loads were followed until week 20 post-infection. Tissue proviral loads were determined as well. The immune response was monitored by measuring FeLV whole virus and p45 antibodies; and feline oncornavirus-associated cell membrane antigen (FOCMA) assay. One cat showed regressive infection (transient antigenemia, persistent provirus-positivity, and seroconversion) with provirus only found in some organs at sacrifice. In 7 of the 20 remaining cats FOCMA assay positivity was the only sign of infection, while all other tests were negative. Overall, the results show that FeLV low dose exposure can result in seroconversion during a presumed abortive infection. Therefore, commonly used detection methods do not detect all FeLV-infected animals, possibly leading to an underestimation of the prevalence of infection.     

The kinetics of feline leukaemia virus shedding in experimentally infected cats are associated with infection outcome

Feline leukaemia virus (FeLV) infection in felids results mainly from oronasal exposure to infectious saliva and nasal secretions, but the potential for viral transmission through faeces and urine has not been completely characterized. In order to assess and compare potential FeLV transmission routes, we determined the viral kinetics in plasma, saliva, faeces and urine during early experimental FeLV infection (up to week 15 post-exposure) in specific pathogen-free cats. In addition to monitoring p27 antigen levels measured by ELISA, we evaluated the presence of infectious particles by cell culture assays and quantified viral RNA loads by a quantitative real-time TaqMan polymerase chain reaction. RNA load was associated with infection outcome (high load-progressive infection; low load-regressive infection) not only in plasma, but also in saliva, faeces and urine. Infectious virus was isolated from the saliva, faeces and urine of infected cats with progressive infection as early as 3-6 weeks post-infection, but usually not in cats with regressive infection. In cats with progressive infection, therefore, not only saliva but also faeces and to some extent urine might represent potential FeLV transmission routes. These results should be taken into account when modelling FeLV-host interactions and assessing FeLV transmission risk. Moreover, during early FeLV infection, detection of viral RNA in saliva may be used as an indicator of recent virus exposure, even in cats without detectable antigenaemia/viraemia. To determine the clinically relevant outcome of FeLV infection in exposed cats, however, p27 antigen levels in the peripheral blood should be measured.     

Soil properties and plant community relationship in a saltmarsh of the Grado and Marano lagoon (northern Italy)

Purpose

The relationship between soil properties and plant communities was investigated in a saltmarsh of the Grado and Marano lagoon (northern Italy), where hydrology and micromorphology strongly influence the features of the ecosystem. A multidisciplinary approach was used to assess the change of soil properties and plant communities in relation to the submergence of soil.

Materials and methods

The plant community and soil profile surveys were both carried out along a transect in six sampling sites of the Gran Chiusa saltmarsh (Grado and Marano lagoon). The morphological and physicochemical parameters of soil profiles were investigated, and soils were classified according to Soil Taxonomy. The concentration of macronutrients in both soils and plants was analysed by inductively coupled plasma-optical emission spectrometry. Cluster and linear discriminant analysis were used to assist the interpretation of the data of plant communities and soil properties, respectively. The bioconcentration factor explored the macronutrient relationship between plant community and soil.

Results and discussion

A high, middle and low zone were identified by clustering the different plant communities along the studied transect. Discriminant analysis showed how the increase in soil submergence supported the accumulation of S and Ca content and depletion of Fe and Na. The development of different plant communities was linked to both soil water saturation and to the capacity of halophytes to tolerate anoxic conditions or salinity, by extrusion or bioconcentration strategies.

Conclusions

This study demonstrates that tide level plays an important role in the pedological development and chemical transformations along a soil hydrosequence. The micromosaic vegetation pattern may therefore represent a useful index of the hydrological and nutritional status of the underlying soils and could be used to predict changes in coastal ecosystems.

     

Osseous sequestration in cattle: 110 cases (1987-1997)

OBJECTIVE: To determine risk factors for development of sequestra in cattle and identify factors associated with a successful outcome. DESIGN: Retrospective study. ANIMALS: 110 cattle. PROCEDURE: Medical records of cattle treated at veterinary teaching hospitals in North America were reviewed. To determine risk factors for osseous sequestration, breed, age, and sex of cattle with osseous sequestration were compared with breed, age, and sex of all other cattle admitted during the study period. RESULTS: 110 cattle were included in the study. Three had 2 sequestra; thus, 113 lesions were identified. Most sequestra were associated with the bones of the extremities, most commonly the third metacarpal or third metatarsal bone. Ninety-two animals were treated surgically (i.e., sequestrectomy), 7 were treated medically, 3 were initially treated medically and were then treated surgically, and 8 were not treated. Follow-up information was available for 65 animals treated surgically and 6 animals treated medically. Fifty-one (78%) animals treated surgically and 5 animals treated medically had a successful outcome. Cattle that were 6 months to 2 years old had a significantly increased risk of developing a sequestrum, compared with cattle < 6 months old. Cattle in which sequestrectomy was performed with the aid of local anesthesia were significantly more likely to undergo 2 or more surgical procedures than were cattle in which sequestrectomy was performed with the aid of general anesthesia. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that sequestrectomy will result in a successful outcome for most cattle with osseous sequestration.     

Growth and mortality of larval anchoveta Engraulis ringens,in northern Chile during winter and their relationship with coastal hydrographic conditions

Larval growth, age, growth effect and instantaneous mortality were estimated in anchoveta, Engraulis ringens, collected biweekly during the austral winter of 2014 in nearshore waters off Bay of Antofagasta (23°41′W–70°30′S), northern Chile. Through measuring standard length (SL) and sagitta microstructure analysis, it was estimated that the growth rate of E. ringens larvae decreased from June (0.85 mm day^?1) to August (0.50 mm day^?1). However, the water temperature was homogeneous during the sampling dates (14.6, 15.2, 14.4, and 14.6°C), suggesting that the decelerating larval growth was not linked to changes in sea temperature. Additionally, larvae with slow growth have larger otoliths compared with conspecifics with fast growth (growth effect). Larval mortality rates tended to decrease until the middle of July (0.18 per day) but increased to 0.25 per day in early August, which coincided with lower food availability (i.e., chlorophyll‐a, 2.7–5.6 mg m^?3) and a high occurrence of smaller larvae (1.58–11.5 mm). Partial least squares analysis indicates low covariance between the biological and oceanographic variables (PLS: 11.71%), suggesting that other factors, such as parental effects, may explain the abrupt decrease in the larval growth rates.     

Presence of Oxytocin,Vasopressin and Atrial Natriuretic Peptide and Their Modification in Rat Hypothalamic Paraventricular Nucleus During Resistance Training

Many studies have demonstrated the physiological effects of oxytocin (OT), atrial natriuretic peptide (ANP) and vasopressin (VP) in the homoeostasis of body fluids during physical exercise. However, a little information is available about the related immunohistochemical changes in hypothalamic magnocellular neurosecretory system during and after the training. The aim of the present work was to study the immunohistochemical changes in OT, ANP and VP levels in the hypothalamic paraventricular nucleus during and after resistance exercise protocol. Three groups of Wistar rats were trained by a rung ladder protocol for 15, 30 and 45 days, respectively; a fourth group was left to rest for 15 days after the training. Finally, four sedentary groups were used as controls. The results show that resistance training induces a significant reduction in the percentage of OT‐positive neurons, compared with sedentary controls. In contrast, this protocol did not induce any change in VP levels, and ANP levels did not change significantly. However, VP increased after the resting period of 15 days. Our work shows that neurons of the paraventricular nucleus are involved in body fluid homoeostasis during and after resistance exercise. The functional significance of these changes in OT and VP levels, during and after the protocol, needs to be further investigated.