Identification and localization of a 94 kDa membrane protein found in Mycoplasma bovoculi strains

Six isolates of Mycoplasma bovoculi obtained from cattle herds with bovine keratoconjunctivitis were analyzed by gel electrophoresis and immunoblotting techniques. All six strains showed similarity in their protein profiles although no two patterns were identical. Antigenic differences between strains were detected in immunoblots reacted with post-exposure calf serum. A common 94 kDa protein band designated p94 was detected in all six strains reacted with monoclonal antibody MA25.5 developed to one of the strains. The p94 was also recognized in these strains by the calf serum. Trypsin treatment of intact mycoplasma cells resulted in the removal of p94 from immunoblots reacted with MA or hyperimmune rabbit serum. Other trypsin-resistant antigens shared between strains or being strain-specific in nature were identified when trypsin-treated mycoplasma cells were reacted with hyperimmune rabbit serum. The p94 antigen was shown to be of mycoplasmal origin by radio-immunoprecipitation using the MA or hyperimmune rabbit serum. These studies identify the presence of a surface antigen (p94) on M. bovoculi membrane in all strains examined that is trypsin sensitive by the use of monoclonal antibody, calf serum and hyperimmune rabbit serum.

Résumé

Six souches de Mycoplasma bovoculi, d'origine bovine (animaux affectés de keratoconjunctivite) ont été analysées par des techniques d'éléctrophorèse et par ‘immunoblotting’. Les six souches ont montré des similitudes dans leurs profils proteiniques. Cependant, aucun profil n'a été identique. Des differences antigéniques entre les souches ont étés detectées suite au traitement des immunoblots avec un sérum bovin infecté par M. bovoculi. L'anticorps monoclonal MA25.5 dirigé contre une des souches a réagi avec une bande proteinique (94 kDa) commune aux six souches. Cette bande a été designee p94. Cette bande p94 a été également identifiée chez tous les souches en utilisant le sérum bovin. Le traitement des mycoplasmes intactes par la trypsine a éliminé la bande p94 des immunoblots: cela a été démontr é par l'usage d'un anticorps monoclonal, ou par un sérum hyperimmun de lapin. D'autres antigènes résistants à la trypsine, communs ou spécifiques aux souches ont été identifiés quand les mycoplasmes traités par le trypsine ont été mélangés au sérum hyperimmun de lapin. La radioimmunoprécipitation par l'anticorps monoclonal ou le sérum hyperimmun de lapin a demontré que l'antigène p94 est d'origine mycoplasmique. Ces études identifient la présence d'un antigène de surface (le p94) localisé sur la membranc de M. bovoculi chez toutes les souches examinées. Cet antigène est sensible à la trypsine. Cela a été montré avec l'anticorps monoclonal, le sérum bovin et le sérum hyperimmune du lapin.     

Behavioral and physiological evaluation of sows raised in outdoors systems in the Brazilian semiarid region

Tropical Animal Health and Production - The objective of this study was to evaluate the biochemical, behavioral, and physiological parameters in pregnant sows of native and improved crossbreeds...     

Serum α-globulin fraction in horses is related to changes in the acute phase proteins

Serum electrophoresis is a useful technique to diagnose some diseases characterized by quantitative and qualitative changes in the serum proteins, such as the acute phase proteins. An increased synthesis of acute phase proteins has been associated with acute infection and inflammation. We wanted to study whether gastrointestinal diseases and disorders of the locomotor system in horses produce enough inflammatory reaction so the acute phase proteins can be detected by electrophoresis in serum. Serum total proteins were calculated, and the serum electrophoresis from 63 horses was performed. The horses were classified in 3 groups: Group I (19 healthy horses), Group II (20 horses suffering from inflammatory disorders of the gastrointestinal tract), and Group III (24 horses with inflammatory disorders of the locomotor system). Results indicated an increase of the acute phase proteins only in patent inflammatory disorders that showed a rapid progression. A global increase in those proteins was observed only for α₁-globulins from Group II.     

Uncertainty assessment of soya bean yield gaps using DSSAT‐CSM‐CROPGRO‐Soybean calibrated by cultivar maturity groups

Soya bean yield gap can be caused by different factors resulting in uncertainties when the objective is to use such information for farm decision‐making and reference yield determination. Thus, this study aimed to quantify the soya bean yield gap for four sites, located in Southern and Midwestern Brazil, as well as the uncertainties of that related to cultivars, sowing dates, soil types and reference yields. The crop simulation model DSSAT‐CSM‐CROPGRO‐Soybean was calibrated for cultivars with similar maturity groups, based on the data obtained from the best farmers at the county level. The yield gap by water deficit (YG_WD) was obtained through the difference between potential and attainable yields, and that one caused by sub‐optimum crop management (YG_CM) by subtracting actual yield of each county, obtained from official statistics between 1989/90 and 2014/15 growing seasons, from the estimated attainable yield. The yield was simulated using four sowing dates, three soil types and two soya bean maturity groups by county. The reference yield uncertainty was quantified using yield reference from crop model and regional winners of the soya bean yield context, conducted by CESB (Brazilian Soybean Strategic Committee), for the growing seasons from 2013/14 to 2015/16. The crop model showed a good agreement between measured and simulated crop development and growth using calibration by maturity group, with low root mean square error (347 kg/ha). Southern sites had a mean YG_WD of 1,047 kg/ha, while in the Midwest, it was lower than 100 kg/ha. The YG_CM was 1,067, 528, 984 and 848 kg/ha, respectively, for Castro, PR, Mamborê, PR, Montividiu, GO and Primavera do Leste, MT, representing the opportunity for yield gain when having the best farmers as reference. The maturity groups, sowing dates and soil types showed to be an important source of uncertainty for yield gap determination, being recommended to investigate the farms in detail for an appropriate quantification. The reference yield showed expressive uncertainties, with some farmers presenting conditions to increase their soya bean yields by more than 3,000 kg/ha, when considering as reference the yields obtained by the winners’ farmers. These results show that uncertainties must be reduced when assessing farm yield gaps, in order to ensure that expected rate of soya bean yield growth could be reached by adopting the same technologies from CESB winners and best farmers in the county as a reference.     

Purification and Characterization of a Novel Alginate Lyase from a Marine Streptomyces Species Isolated from Seaweed

Alginate, a natural polysaccharide derived from brown seaweed, is finding multiple applications in biomedicine via its transformation through chemical, physical, and, increasingly, enzymatic processes. In this study a novel alginate lyase, AlyDS44, was purified and characterized from a marine actinobacterium, Streptomyces luridiscabiei, which was isolated from decomposing seaweed. The purified enzyme had a specific activity of 108.6 U/mg, with a molecular weight of 28.6 kDa, and was composed of 260 amino acid residues. AlyDS44 is a bifunctional alginate lyase, active on both polyguluronate and polymannuronate, though it preferentially degrades polyguluronate. The optimal pH of this enzyme is 8.5 and the optimal temperature is 45 °C. It is a salt-tolerant alginate lyase with an optimal activity at 0.6 M NaCl. Metal ions Mn²⁺, Co²⁺, and Fe²⁺ increased the alginate degrading activity, but it was inhibited in the presence of Zn²⁺ and Cu²⁺. The highly conserved regions of its amino acid sequences indicated that AlyDS44 belongs to the polysaccharide lyase family 7. The main breakdown products of the enzyme on alginate were disaccharides, trisaccharides, and tetrasaccharides, which demonstrated that this enzyme acted as an endo-type alginate lyase. AlyDS44 is a novel enzyme, with the potential for efficient production of alginate oligosaccharides with low degrees of polymerization.     

Sodicity and salinity in a Brazilian Oxisol cultivated with sugarcane irrigated with wastewater

Changes in soil sodicity-salinity parameters are one of the most characteristic alterations after treated sewage effluent (TSE) irrigation in agro-systems. Considering the importance of these parameters for agricultural management, as well as the economical value of sugarcane for Brazil, the present study aimed at evaluating effects on soil sodicity and salinity under tropical conditions over 16 months of TSE irrigation in a sugarcane plantation at Lins, São Paulo State, Brazil. Soil samplings were carried out in February 2005 (before planting), December 2005 (after 8 months of TSE irrigation) and September 2006 (after 16 months of TSE irrigation) following a complete block design with four treatments and four replicates. Treatments consisted of: (i) control, without TSE irrigation; (ii) T100, T150 and T200, with TSE irrigation supplying 100% (0% surplus, total of 2524 mm), 150% (50% surplus, total of 3832 mm) and 200% (100% surplus, total of 5092 mm) of crop water demand, respectively. Compared to initial soil conditions, at the end of the experiment increases of exchangeable sodium (from 2.4 to 5.9 mmol_c kg⁻¹), exchangeable sodium percentage (ESP) (from 8 to 18%), soluble Na (from 1.4 to 4.7 mmol L⁻¹) and sodium adsorption ratio (SAR) of soil solution (from 3.6 to 12.6 (mmol L⁻¹)^0.5) were found in the soil profile (0-100 cm) as an average for the irrigated plots due to high SAR of TSE. Associated with the increments were mostly significant increases in clay dispersion rates at depths 0-10, 10-20 and 20-40 cm. Electrical conductivity (EC) of soil solution increased during the TSE irrigation period whereas at the end of the experiment, after short term discontinuation of irrigation and harvest, EC in the topsoil (0-10 and 10-20 cm) decreased compared to the previous samplings. Moreover, despite increasing sodicity over time mainly insignificant differences within the different irrigated treatments were found in December 2005 and September 2006. This suggests that independent of varying irrigation amounts the increasing soil sodicity over time were rather caused by the continuous use of TSE than by its quantity applied. Moreover, also plant productivity showed no significant differences within the TSE irrigated plots. The study indicates that monitoring as well as remediation of soil after TSE irrigation is required for a sustainable TSE use in order to maintain agricultural quality parameters.     

Outbreak of shoot blight and dieback of Eucalyptus spp., caused by Pseudoplagiostoma eucalypti in Brazil

An outbreak of a new and severe disease was observed in Eucalyptus plantations of Bahia state, Brazil. An Ascomycota fungus has been frequently associated with the main symptoms of the disease namely leaf spot, branch cankers, shoot blight, defoliation, and dieback. Based on morphological characteristics, phylogenetic analysis (ITS and TEF-1α genes), and pathogenicity test on Eucalyptus plants, Pseudoplagiostoma eucalypti was identified as the causal agent of the disease. Although P. eucalytpi has been known from in Brazil since 1998, this is the first report of it causing severe disease and die-back on Eucalyptus spp. and we also record new symptoms associated with the pathogen.     

Inheritance of race 1.2 Fusarium wilt resistance in four melon cultivars

The resistance to Fusarium oxysporum f.sp. melonis (Fom) race 1.2 has been studied in melons, such as the Portuguese accession ‘BG-5384’ and in the Japanese ‘Shiro Uri Okayama’, ‘Kogane Nashi Makuwa’, and ‘C-211’, since a good characterization of the resistance is necessary before its introgression into commercial varieties. These four melon accessions showed a high level of resistance to races 0, 1, and 2 of Fom, indicating that the partial resistance to the race 1.2 previously detected may not have been race specific. To determine the mode of inheritance of the resistance to Fom race 1.2, the F₁, F₂, BCP_R, and BCP_S generations from the crosses between the four resistant accessions above and ‘Piel de Sapo’, a Fom race 1.2 susceptible melon, were developed. They were subsequently inoculated with two Fom isolates, one from the pathotype 1.2Y and the other from the pathotype 1.2W. The area under the disease progress curve was determined for each inoculated plant, and the data were analyzed. We show that the resistance seen in these accessions is polygenically inherited with a complex genetic control because many epistatic interactions were detected. The three epistatic effects; additivity × additivity, dominance × dominance, and dominance × additivity are present and significant, with differing magnitudes from one cross to the next. The relatively low heritabilities, and these epistatic effects make difficult the improvement of the resistance, from these sources, through a standard selection procedure.     

Evaluation of analgesic and physiologic effects of epidural morphine administered at a thoracic or lumbar level in dogs undergoing thoracotomy

ObjectiveTo evaluate the analgesic and physiological effects of epidural morphine administered at the sixth and seventh lumbar or the fifth and sixth thoracic vertebrae in dogs undergoing thoracotomy.Study designProspective, randomized, blinded trial.AnimalsFourteen mixed-breed dogs, weighing 8.6 ± 1.4 kg.MethodsThe animals received acepromazine (0.1 mg kg^?1) IM and anesthesia was induced with propofol (4 mg kg^?1) IV. The lumbosacral space was punctured and an epidural catheter was inserted up to the region between the sixth and seventh lumbar vertebrae (L, n = 6) or up to the fifth or sixth intercostal space (T, n = 8). The dogs were allowed to recover and after radiographic confirmation of correct catheter position, anesthesia was reinduced with propofol IV and maintained with 1.7% isoflurane. Following stabilization of monitored parameters, animals received morphine (0.1 mg kg^?1) diluted in 0.9% NaCl to a final volume of 0.25 mL kg^?1 via the epidural catheter, and after 40 minutes, thoracotomy was initiated. Heart rate and rhythm, systolic, mean and diastolic arterial pressures, respiratory rate, arterial hemoglobin oxygen saturation, partial pressure of expired CO₂ and body temperature were measured immediately before the epidural administration of morphine (0 minute) and every 10 minutes during the anesthetic period. The Melbourne pain scale and the visual analog scale were used to assess post-operative pain. The evaluation began 3 hours after the epidural administration of morphine and occurred each hour until rescue analgesia.ResultsThere were no important variations in the physiological parameters during the anesthetic period. The post-operative analgesic period differed between the groups, being longer in T (9.9 ± 1.6 hours) compared with L (5.8 ± 0.8 hours).ConclusionsThe use of morphine, at a volume of 0.25 mL kg^?1, administered epidurally over the thoracic vertebrae provided longer lasting analgesia than when deposited over the lumbar vertebrae.Clinical relevanceThe deposition of epidural morphine provided longer lasting analgesia when administered near to the innervation of the injured tissue without increasing side effects.