Serobiotypes and virulence genes of Escherichia coli strains isolated from diarrheic and healthy rabbits in Brazil

A total of 178 Escherichia coli isolates from diarrheic and healthy rabbits in the S?o Paulo State (Brazil) were serobiotyped and investigated by PCR for the presence of virulence genes. Among the 90 (50.6%) isolates which possessed the eae gene, 74 were from diarrheic animals and all but one encoded intimin beta. Sixty five (72.2%) of the eae+ isolates had insertion of the locus of enterocyte effacement locus in the pheU locus, 11 (12.2%) in the selC and 14 (15.6%) did not insert in either of these loci. All isolates were negative for genes of the E. coli enterotoxins, Stx1, Stx2, CNF1, CNF2 and EHEC hemolysin. The O132:H2 serotype was dominant, being present in 63 isolates (70%) of the 90 eae+ isolates, and 57 of the 63 isolates of this serotype belonged to biotype 30. PCR detected the gene for AF/R2 fimbriae in 75 (83.3%) of the 90 eae+ isolates. Adherence to HeLa cells was best detected following 6h incubation and a positive fluorescence actin staining (FAS) test was given by 52 isolates. These data show that isolates of E. coli associated with diarrhea in rabbits in Brazil possess the genotype and phenotype typically associated with rabbit enteropathogenic E. coli (EPEC). We conclude that EPEC that possess the eae gene are a common cause of diarrhea in Brazilian rabbit farms and that the pathogenic eae+ AF/R2+ isolates of O132:H2:B30 serobiotype are especially predominant.     

TRAP molecular markers as a system for saturation of the genetic map of durum wheat

Target region amplification polymorphism (TRAP) is a relatively new PCR-based technique that detects large numbers of loci in a single reaction without extensive pre-PCR processing of samples. The aim of this study was to integrate TRAP markers in an EST-derived SSR linkage map of a RIL mapping population from the cross of the durum wheat cultivars Ciccio and Svevo, for a more general purpose of establishing a high-throughput system for genetic map saturation. Primer combinations producing PCR products with at least 4–5 polymorphic bands were selected and analyzed across the mapping population. The PCR reactions produced a total of 2,881 fragments with an average of 52 peaks per reaction. A total of 142 new TRAP markers were mapped and found to be randomly distributed in the genome. The total length of the map was 2,043.0 cM, with an average chromosome length of 145.9 cM. Homoeologous group one had the highest number of TRAP markers (38 loci) and the longest map length (407.9 cM) for a total of 87 markers, while the homoeologous group five had the lowest TRAP marker number (5 loci) and the shortest map length (232.5 cM). The distribution of markers among the seven homoeologous groups was random. The results indicate that TRAP is highly efficient in genetic mapping, generating a large number of markers scattered across the genome. This closes many existing gaps in marker coverage and may join otherwise separate linkage groups.     

Early evaluation of popcorn inbred lines for phosphorus use efficiency

Phosphorus use efficiency (PUE) evaluation based on shoot dry weight in the early stages of plant development can be a useful measure to identify genotypes that are superior in phosphorus (P) use. The objectives of this study were to (i) identify inbred lines that are efficient and inefficient in P use, (ii) assess the genetic diversity among the inbred lines based on traits related to PUE, (iii) investigate the relationship among traits related to PUE and its components and (iv) estimate the expected responses to selection for increasing PUE. Twenty‐three S₆ popcorn inbred lines were evaluated under high and low P, and several traits related to PUE and its components were measured. Efficient and inefficient inbred lines under each P level were identified, and the genetic diversity assessment grouped these inbred lines in opposite clusters. Phosphorus acquisition efficiency (PAE) was the most important component of PUE in the early stages of plant development, and the direct selection for PAE seems to be the best method for increasing PUE in these stages.     

Morphometric analyses of the body and the branches of the normal third interosseous muscle (suspensory ligament) in Standardbreds

The third interosseous muscle (suspensory ligament, TIOM) is composed of connective tissue (CT) with a variable proportion of muscle (MT) and adipose tissue (AT). The aim of our study is to quantify the CT, MT and AT within the body and the branches of right thoracic and pelvic limbs TIOM in sound horses to determine whether there are differences in CT, MT and AT between age, sex, limbs and levels. Right limbs from 11 sound horses were collected. Samples from 6 levels of the TIOM were embedded in paraffin or in Tissue‐Tek^®. Most of the paraffin sections were shredded. Using the cryosection, some artefacts appeared. Cryoprotection was carried out, which produced the best results. Hematoxylin–phloxine–saffron and Hematoxylin–eosin gave a good contrast of colours between the tissues observed allowing the use of an image analysis programme to calculate percentage of each tissue within the TIOM. The percentage of MT and AT decreased significantly (P < 0.0001), whereas the percentage of CT increased significantly (P < 0.0001) with age and when descending from the proximal to the distal level of the TIOM. The percentage of MT was significantly higher (P < 0.0001) in females than males, while the percentage of CT was significantly higher (P < 0.0001) in males than females. The percentage of AT was significantly higher (P = 0.0278) in pelvic limbs than in thoracic limbs. These results confirm the variation in tissue composition within the TIOM of sound horses.     

Effects of Pentoxifylline on Equine Epididymal Sperm

This study evaluated whether pentoxifylline (PTX) present in the flushing extender influenced the function of equine epididymal spermatozoa after recovery and after thawing. For this experiment, 58 testicles from 29 Brazilian Jumping Horses were used. Cauda epididymides of each stallion were separated and flushed with a skim milk extender, with or without 7.18 mM PTX and then subjected to the freezing process. Samples flushed with the extender containing PTX showed a significant increase in total motility, progressive motility, straight line velocity, curvilinear velocity, and percentage of rapid sperm immediately after the recovery of epididymal sperm and after 15 minutes of incubation at 37°C (P < .05). However, the presence of PTX in the flushing extender did not affect the post-thaw motility parameters or plasma membrane integrity (P > .05). The results of this study showed that the PTX present in the flushing extender improved motility parameters of recently recovered epididymal sperm and had no deleterious effects on plasma membrane integrity and freezability of equine epididymal sperm.     

Presence of Leptospiral DNA in Semen Suggests Venereal Transmission in Horses

The purpose of the present study was to detect leptospiral DNA by PCR in semen and urine samples of stallions to test for venereal transmission in horses. A total of 10 stallions from four herds were studied, and sampling was conducted in semen and urine for culture and PCR and serum for serology. From the 10 serum samples tested, 6 (60%) were seroreactive. No pure culture was obtained, but leptospiral DNA was detected by PCR in 50% of the semen samples and 30% of urine samples. The present study aimed to detect leptospiral DNA by PCR in semen and urine samples of stallions to test for venereal transmission in horses. Based on these findings, we suggest that there is potential transmission of leptospirosis in horses by sexual transmission.     

The relationships between scrotal surface temperature,age and sperm quality in stallions

In horses, spermatogenesis normally occurs at an average intratesticular temperature of 35 °C; therefore, mechanisms for testicular thermoregulation are essential. Measuring the scrotal surface temperature by thermography is one of the methodologies used to evaluate the effectiveness of testicular thermoregulation. The objective of this study was to determine the relationship between the control of scrotal surface temperature and sperm quality in horses of different ages. In total, 24 Quarter Horse stallions were divided into three groups: YS (young stallions), AS (adult stallions) and OS (old stallions). Initially, we calculated the testicular volume (TV) and evaluated various aspects of the semen (sperm kinetics, plasma membrane integrity and sperm morphology) for all the animals. We also evaluated rectal temperature (RT), body surface temperature (BST,) and average scrotal surface temperature in the testicular region (SST) before (M0) and after sun exposure (M1). Differences were observed (p<0.05) between the RT and BST before and after sun exposure in all three groups. However, there were no differences (p>0.05) in the SST values at these two time points, thus demonstrating the efficiency of the mechanisms for testicular thermoregulation. The SST was similar (p>0.05) among all three groups. Based on these results, we conclude that fertile stallions of different age groups are able to maintain SST and measuring the heat radiating from the scrotum using a digital infrared thermographer. We can also conclude that measuring the heat radiating from the scrotum using a digital infrared thermographer is a practical and efficient tool for monitoring SST in horses.     

Landscape pattern changes over 25 years across a hotspot zone in southern Brazil

Forest fragmentation caused by human activities has many implications for natural landscapes, such as habitat reduction and the loss of biodiversity. This study investigated the temporal fragmentation process of forest remnants in a strongly agro-industrialised region in southern Brazil over 25 years. The studied watershed area hosts two important typologies of the Atlantic Forest biome as well as Quaternary remnants of the Brazilian Savanna biome, which are considered hotspots of biodiversity and reflect the intense process of forest fragmentation caused by Brazilian urban and agro-industrial development. Thus, studies encompassing multitemporal scales are paramount to understanding changes in forest patterns and are fundamental for trend predictions of landscape dynamics. To perform the calculation of the mean normalised difference vegetation index, Landsat satellite images from 1991 to 2016 were processed using SPRING® software. Subsequently, FRAGSTATS® software was used to calculate landscape metrics. A reduction in the number of forest fragments since 1991 was observed, with a maximum amount of 5 243 fragments in 1993 that declined to 4 015 fragments in 2016. Although the number of fragments in the watershed decreased, the mean area increased by 72.9% and the mean of the shape index increased from 1.3 in 1991 to 1.5 in 2016. In addition, there was a 64.7% increase in the edge density and a reduction of 35.6 m in the isolation between the nearest neighbours. The degree of isolation of the fragments underwent a process of expansion and reduction when compared to 1991, presenting results that support the hypothesis that the Atlantic Forest is in a process of stabilisation and forest restoration.     

CO2 fertilization does not affect biomass production and nutritive value of a C4 tropical grass in short timeframe

Increased atmospheric carbon dioxide (CO₂) is a consequence of recent anthropogenic environmental changes, and few studies have evaluated its effects on tropical grasses used in Brazilian pastures, the main feed source for major part of ruminant livestock. This study evaluated forage production, chemical composition, in vitro total gas production and organic matter degradability of Brachiaria brizantha under contrasting CO₂ atmospheric conditions in a free air carbon dioxide enrichment (FACE) facility. The forage plants were sown in each of the 12 octagonal rings of the FACE facility: six under ambient atmospheric CO₂ concentration of approximately 390 μmol/mol, hereafter referred to as control (CON) plots, and other six rings enriched with pure CO₂ flux to achieve a target CO₂ concentration of 550 μmol/mol, hereafter called elevated CO₂ (eCO₂) plots. Soil samples were collected to determine carbon and nitrogen concentrations. After seventy days of sowing, a standardization cutting was performed and then at regular intervals of 21 days the forage was harvested (ten harvest dates) and forwarded to laboratorial analyses. Forage above‐ground biomass production (dry matter (DM): 6,143 vs. 6,554 kg/ha), as well as morphological characteristics (leaves: 71% vs. 68%; stem: 28% vs. 31%), chemical composition (crude protein: 162.9 vs. 161.8; neutral detergent fibre: 663.8 vs. 664.3; acid detergent fibre: 369.5 vs. 381; lignin: 60.1 vs. 64.1 g/kg DM; total C: 45.9 vs. 45.9; total N: 2.8 vs. 2.8; total S: 0.2% vs. 0.2%), organic matter in vitro degradability (573.5 vs. 585.3 g/kg), methane (5.7 vs. 4.3 ml/g DM) and total gas (128.3 vs. 94.5 ml/g DM) production did not differ significantly between CON and eCO₂ treatments (p > .05). The results indicated that at least under short‐term enrichment, B. brizantha was not affected by eCO₂.     

The use of infrared spectroscopy and machine learning tools for detection of Meloidogyne infestations

Plant parasitic nematodes are generally soilborne pathogens that attack plants and cause economic losses in many crops. The infested plants show nonspecific symptoms or, often, are symptomless; therefore, diagnosis is performed by taking soil and root tissue samples. Here, we show that a combination of different infrared spectra analysis and machine learning algorithms can be used to detect plant parasitic nematode infestations before symptoms become visible, using leaves instead of roots and soil as samples. We found that tomato and guava plants infested with Meloidogyne enterorlobii produced different spectral patterns compared to uninfested plants. Using partial spectra from 1,450 to 900/cm as the "fingerprint region", principal component analyses indicated that after 5 (tomatoes) or 8 weeks (guava), plants with no visible symptoms of infestations were positively diagnosed. To improve the early detection response, we used machine learning modelling. A support vector machine (SVM) was used to obtain more robust, accurate models. The SVM model contained 34 support vectors, 17 for each level. The overall performance of the model was >97% and the total accuracy was significantly higher, demonstrating the absence of chance prediction. The best prediction of infestation was obtained at the second and fourth weeks for tomatoes and guavas, respectively, reducing the diagnostic time by half. The combined application of these techniques reduces the processing time from field to laboratory and shows enormous advantages by avoiding root and soil sampling.