Identification of viral pathogens in aborted fetuses and stillborn piglets from cases of swine reproductive failure in Spain

The objective of the present study was to determine the presence of recognised abortifacient viruses such as porcine reproductive and respiratory virus (PRRSV), Aujeszky's disease virus (ADV), porcine parvovirus (PPV) and porcine circovirus type 2 (PCV2), in tissues from aborted fetuses and stillborn neonates in cases of late reproductive failure in swine. A total of 293 specimens (fetuses aborted in the last third of gestation and stillborn piglets) from 100 different cases of late-term abortions and premature farrowing from 15 different Spanish provinces were studied. PRRSV was detected in 9/100 cases by RT-PCR. Only 1/100 cases analysed (corresponding to a late-term aborted fetus with a negative PRRSV RT-PCR result) was positive for PCV2 by PCR. Neither ADV (monitored by viral isolation plus antigen detection) nor PPV (monitored by ELISA antigen capture test) infection was identified. The results suggest that PRRSV is one of the most important infectious agents, if not the most relevant one, associated with fetal infection leading to abortion or premature farrowing in Spain. Moreover, other viral pathogens such as ADV, PPV and PCV2 seem to have a minor impact on reproductive disease.     

Description of the infection status in a Norwegian cattle herd naturally infected by Mycobacterium avium subsp. paratuberculosis

The Norwegian surveillance and control programme for paratuberculosis revealed 8 seroreactors in a single dairy cattle herd that had no clinical signs of Mycobacterium avium subsp. paratuberculosis (M. a. paratuberculosis) infection. Paratuberculosis had been a clinical problem in goats several years previously in this herd. All 45 cattle were culled and a thorough investigation of the infection status was conducted by the use of interferon-gamma (IFN-gamma) immunoassay, measurement of antibodies, and pathological and bacteriological examination. In the IFN-gamma immunoassay, 9 animals gave positive results, and 13 were weakly positive, while 19 animals were negative. In the serological test, 10 animals showed positive reactions, and 5 were doubtful, while 30 animals gave negative reactions. There appeared to be a weak trend toward younger animals having raised IFN-gamma and older animals having raised serological tests. Histopathological lesions compatible with paratuberculosis were diagnosed in 4 animals aged between 4 and 9 years. Three of these animals had positive serological reaction and one animal gave also positive results in the IFN-gamma immunoassay. Infection was confirmed by isolation of M. a. paratuberculosis from 2 of these 4 animals. One single bacterial isolate examined by restriction fragment length polymorphism (RFLP) had the same profile, B-C1, as a strain that had been isolated from a goat at the same farm several years previously. Despite many animals being positive in one or both of the immunological tests, indicative of a heavily infected herd, none of the animals showed clinical signs and only one cow was shown to be shedding bacteria. A cross-reaction with other mycobacteria might have caused some of the immunoreactions in these animals. It is also possible that the Norwegian red cattle breed is resistant to clinical infection with M. a. paratuberculosis.     

Pharmacokinetics of azithromycin after i.v. and i.m. administration to sheep

The pharmacokinetics (PK) of azithromycin after i.v. and i.m. injection at a single dosage of 20 mg/kg bodyweight was studied in sheep. Blood samples were collected from the jugular vein until 120 h after dosing for both routes. Plasma concentrations of azithromycin were determined by bioassay. The plasma concentration-time data of azithromycin best fitted a three-compartment model after i.v. administration and a two-compartment model with first-order absorption after i.m. administration. The elimination half-life (t(1/2lambdaz)) was 47.70 +/- 7.49 h after i.v. administration and 61.29 +/- 13.86 h after i.m. administration. Clearance value after i.v. dosing was 0.52 +/- 0.08 L/kg.h. After i.m. administration a peak azithromycin concentration (C(max)) of 1.26 +/- 0.19 mg/L was achieved at 1.24 +/- 0.31 h (t(max)). Area under the curve (AUC) were 38.85 +/- 5.83 mg.h/L and 36.03 +/- 1.52 mg.h/L after i.v. and i.m. administration respectively. Bioavailability obtained after i.m. administration was 94.08 +/- 11.56%. The high tolerability of this i.m. preparation and the favourable PK behaviour such as the long half-life and high bioavailability make azithromycin likely to be effective in sheep.     

Preliminary studies of serum acute-phase protein concentrations in hematologic and neoplastic diseases of the dog

Serum concentrations of acute-phase proteins (APPs): haptoglobin (Hp), ceruloplasmin (Cp), serum amyloid A (SAA), and C-reactive protein (CRP) were determined in healthy dogs (n = 15) and dogs with different diseases grouped as acute inflammation (I, n = 12), hematologic neoplasias (HT, including leukemia and lymphoma, n = 16), nonhematologic neoplasias (NHT, including epithelial, mesenchymal, and mixed, n = 20), and autoimmune hemolytic anemia (AIHA, n = 8). SAA and CRP were analyzed using commercially available enzyme-linked immunosorbent assay (ELISA) kits, and Hp and Cp were measured using colorimetric methods, all previously validated for use in dogs. Increased concentrations of all APPs were observed in all groups of diseased dogs, but statistical significance only was observed with Hp (I, P < .001; HT, P < .05), Cp (I, P < .05; AIHA, P < .01), and CRP (I, P < .001; HT, P < .001; AIHA, CRP P < .05). High variability in individual APPs within each group of diseases was found with no significant differences between leukemia and lymphoma as well as among different types of neoplasia. The AIHA group had smaller increases in Hp, SAA, and CRP but higher concentrations of Cp. When follow-up of individual cases was possible, a decrease in APPs generally was found in cases with favorable outcome. The results of this study suggest that neoplasia and hematologic diseases such as AIHA should be considered as possible causes of mild increases in APPs in dogs. Measurement of APPs may be helpful to assess clinical evolution and monitor treatment of these processes.     

Overland water and salt flows in a set of rice paddies

Cultivation of paddy rice in semiarid areas of the world faces problems related to water scarcity. This paper aims at characterizing water use in a set of paddies located in the central Ebro basin of Spain using experimentation and computer simulation. A commercial field with six interconnected paddies, with a total area of 5.31 ha, was instrumented to measure discharge and water quality at the inflow and at the runoff outlet. The soil was classified as a Typic Calcixerept, and was characterized by a mild salinity (2.5 dS m⁻¹) and an infiltration rate of 5.8 mm day⁻¹. The evolution of flow depth at all paddies was recorded. Data from the 2002 rice-growing season was elaborated using a mass balance approach to estimate the infiltration rate and the evolution of discharge between paddies. Seasonal crop evapotranspiration, estimated with the surface renewal method, was 731 mm (5.1 mm day⁻¹), very similar to that of other summer cereals grown in the area, like corn. The irrigation input was 1874 mm, deep percolation was 830 mm and surface runoff was 372 mm. Irrigation efficiency was estimated as 41%. The quality of surface runoff water was slightly degraded due to evapoconcentration and to the contact with the soil. During the period 2001–2003, the electrical conductivity of surface runoff water was 54% higher than that of irrigation water. However, the runoff water was suitable for irrigation. A mechanistic mass balance model of inter-paddy water flow permitted to conclude that improvements in irrigation efficiency cannot be easily obtained in the experimental conditions. Since deep percolation losses more than double surface runoff losses, a reduction in irrigation discharge would not have much room for efficiency improvement. Simulations also showed that rice irrigation performance was not negatively affected by the fluctuating inflow hydrograph. These hydrographs are typical of turnouts located at the tail end of tertiary irrigation ditches. In fact, these are the sites where rice has been historically cultivated in the study area, since local soils are often saline-sodic and can only grow paddy rice taking advantage of the low salinity of the irrigation water. The low infiltration rate characteristic of these saline-sodic soils (an experimental value of 3.2 mm day⁻¹ was obtained) combined with a reduced irrigation discharge resulted in a simulated irrigation efficiency of 60%. Paddy rice irrigation efficiency can attain reasonable values in the local saline-sodic soils, where the infiltration rate is clearly smaller than the average daily rice evapotranspiration.     

Determination of the crop coefficient for grafted ‘Tahiti’ lime trees and soil evaporation coefficient of Rhodic Kandiudalf clay soil in Sao Paulo,Brazil

The expansion of permanent trickle irrigation systems in Sao Paulo (Brazil) citrus has changed the focus of irrigation scheduling from determining irrigation timing to quantifying irrigation amounts. The water requirements of citrus orchards are difficult to estimate, since they are influenced by heterogeneous factors such as age, planting density and irrigation system. In this study, we estimated the water requirements of young ‘Tahiti’ lime orchards, considering the independent contributions from soil evaporation and crop transpiration by splitting the crop coefficient (Kc = ETc/ETo) into two separate coefficients; Ke, a soil evaporation coefficient and Kcb, a crop transpiration coefficient. Hence, the water requirement in young ‘Tahiti’ lime (ET_y) is ET_y = (Ke + Kcb) · ETo, where ETo is the reference crop evapotranspiration. Mature tree water requirement (ET_m) is ET_m = Kcb · ETo, assuming no soil water evaporation. Two lysimeters were used; one was 1.6 m in diameter and 0.7 m deep, and the other was 2.7 m in diameter and 0.8-m deep. The first one was used to calculate evaporation and the second one was used for transpiration. ETo was estimated by the Penman–Monteith method (FAO-56). The measurements were conducted during a period between August 2002 and April 2005 in Piracicaba, Sao Paulo state, Brazil. The lysimeters were installed at the center of a 1.0-ha plot planted with ‘Tahiti’ lime trees grafted on ‘Swingle’ citrumelo rootstock. The trees were 1-year old at planting, spaced 7 × 4 m, and were irrigated by a drip irrigation system. During the study period, Kc varied between 0.6 and 1.22, and Kcb varied between 0.4 and 1.0. The results suggested that for young lime trees, the volume of water per tree calculated by Ke + Kcb is about 80% higher than the volume calculated using Kc. For mature trees, the volume of water per tree calculated using just Kcb can be 10% less than using Kc. The independent influence of soil evaporation and transpiration is important to better understand the water consumption of young lime trees during growth compared to mature lime trees.     

Dietary Photoprotective Compounds Ameliorate UV Tolerance in Shrimp (Pleoticus muelleri) through Induction of Antioxidant Activity

The aim of this study was to determine the effects of ultraviolet radiation (UVR) on bioaccumulation of UV‐absorbing compounds acquired through the diet, in larvae and postlarvae of Pleoticus muelleri, and to assess tissue antioxidant activity, survival, and development. Mysis stage I were exposed to two artificial radiation treatments: M‐PAR (photosynthetically active radiation, range = 400–700 nm) and M‐PAR + UVR (280–700 nm). The experimental larvae received a mixed dietary treatment of Artemia persimilis and the microalga Pavlova lutheri, reared under two radiation regimes: PAR (D‐PAR) and PAR + UVR (D‐PAR + UVR). Shrimp from all treatments reached 8 d postlarval stage (PL8), except those under M‐PAR + UVR treatment fed the D‐PAR‐cultured algae, which had 0% survival. Larvae in M‐PAR + UVR and M‐PAR treatments fed with D‐PAR + UVR diet presented the highest survival rates (70 and 75%, respectively), with 37 and 41% increase in PL size. UV‐absorbing compounds were detected in microalgae and PL subject to PAR + UVR treatments. Antioxidant activity, quantified by measuring the free‐radical 1,1‐diphenyl‐2‐picrylhydrazyl in homogenates of PL8, decayed drastically under radiation treatment M‐PAR + UVR fed with algae of the D‐PAR + UVR treatment. It is concluded that the bioaccumulation of UV‐absorbing compounds and the highest antioxidant activity in PL could improve the biochemical and photophysiological responses of shrimp under UVR stress.     

Evaluation of the API 50CH and API ZYM systems for rapid characterization of Clavibacter michiganensis subsp. sepedonicus, causal agent of potato ring rot

API 50CH and API ZYM systems were used to characterize fifty-three strains of Clavibacter michiganensis subsp. sepedonicus from different geographic locations and several reference strains of the same and different species, including other potato pathogens. Clavibacter michiganensis subsp. sepedonicus strains displayed a high level of homogeneity, both in carbohydrate utilization and in enzymatic activity. Using API 50CH and API ZYM it was possible to differentiate C. michiganensis subsp. sepedonicus strains from the remaining taxa analysed in this study, which included representative strains of the other subspecies of C. michiganensis as well as other bacterial pathogens affecting potatoes. Therefore, these systems could be used as an effective method to characterize C. michiganensis subsp. sepedonicus. Such a procedure would constitute an alternative system to the conventional nutritional and physiological identification tests currently included in the official methods employed in the European Union to detect and identify this bacterium. The results obtained with the API systems agreed with the current taxonomic classification of C. michiganensis, clearly separating sepedonicus from the other subspecies belonging to this species.     

Biological control of botrytis bunch rot in organic wine grapes with the yeast antagonist Candida sake CPA‐1

The aim of this research was to confirm the efficacy of the yeast antagonist Candida sake CPA‐1 in suppressing botrytis bunch rot development, in an organic vineyard under Mediterranean conditions for two seasons, and compare its performance with that of two biologically based products currently registered for botrytis bunch rot control in New Zealand. In 2009, treatments applied were: commercial formulations of Ulocladium oudemansii (BOTRY‐Zen^®) and chitosan (ARMOUR‐Zen^®), C. sake CPA‐1 combined with the fatty acid‐based additive Fungicover^® and combinations of these products. All treatments were applied six times between early flowering and harvest and compared with an unsprayed control. In 2010, the treatments focused on C. sake and Fungicover and the number of applications was reduced from six to four. The population dynamics of U. oudemansii and C. sake were measured and wine quality tests were carried out in both seasons. Disease control achieved by C. sake treatments in 2009 were comparable to those achieved by BOTRY‐Zen and ARMOUR‐Zen. Applications of C. sake plus Fungicover between flowering and harvest significantly (P < 0·05) reduced botrytis bunch rot incidence and severity by 64% and 90%, respectively, compared with the untreated control in 2009, and by 67% and 89%, respectively, in 2010. Treatments did not adversely affect wine quality parameters after treated grapes were processed. Candida sake consistently provided effective control of botrytis bunch rot in grapes under different meteorological and disease pressure conditions, thereby improving its potential for future commercial applications.     

Relationships between agronomic factors and epidemics of Phytophthora branch canker of citrus in southwestern Spain

Phytophthora branch canker, caused by Phytophthora citrophthora, has been an increasing problem in clementine (Citrus reticulata) production in Spain during last years. The disease was particularly severe in the new citrus-growing areas of the southwestern coastal areas in Huelva Province. Recent studies revealed that disease emergence was not related to either genetic drift or host specificity changes in P. citrophthora population. Therefore, the possible association of agronomic factors with the disease was investigated. A total of 110 orchards were selected arbitrarily from the main citrus-growing areas in Huelva Province. The presence of branch cankers together with agronomic factors including soils, cultivars, rootstocks, irrigation, pruning, techniques to improve fruit production, fungicide treatments, presence of brown rot of fruit and frost damage were recorded. Logistic regression analysis was used to detect correlations between the agronomic factors studied and disease prevalence. Phytophthora branch canker was significantly associated with mature clementine orchards. Sweet orange and hybrid cultivars as well as young clementine orchards were less affected by the disease. Although disease was less frequent in Salorthid soils, alternative high resolution procedures are required to draw conclusions about the effect of soil properties on disease prevalence. As in other Phytophthora-induced diseases, soil flooding during the rainy season was correlated positively with the prevalence of branch cankers. Improving fruit production by branch scoring showed a strong positive correlation with Phytophthora branch canker. This is the first time that girdling has been associated with Phytophthora disease epidemics on a fruit tree crop, but further research is needed to determine the cause of this relationship. Cultural practices including pruning, regulated deficit irrigation, additional phosphonate sprays, and abiotic and disease factors such as frost damage and presence of brown rot of fruit were not significantly correlated with disease prevalence.