Detection of the Defoliating Pathotype of Verticillium dahliae in Infected Olive Plants by Nested PCR

Spread of Verticillium wilt into newly established olive orchards in Andalucía, southern Spain, has caused concern in the olive industry in the region. This spread may result from use of Verticillium dahliae-infected planting material, which can extend distribution of the highly virulent, defoliating (D) pathotype of V. dahliae to new areas. In this study, a molecular diagnostic method for the early in planta detection of D V. dahliae was developed, aimed especially at nursery-produced olive plants. For this purpose, new primers for nested PCR were designed by sequencing a 992-bp RAPD marker of the D pathotype. The use of the specific primers and different nested-PCR protocols allowed the detection of V. dahliae pathotype D DNA in infected root and stem tissues of young olive plants. Detection of the pathogen was effective from the very earliest moments following inoculation of olive plants with a V. dahliae pathotype D conidia suspension as well as in inoculated, though symptomless, plants.     

Effect of cyclodextrin diallyl maleate on methane production, ruminal fermentation and microbes in vitro and in vivo

Effects of β‐cyclodextrin diallyl maleate (CD‐M) on methane production, ruminal fermentation and digestibility were studied both in vitro and in vivo. In in vitro study, diluted ruminal fluid (30 mL) was incubated anaerobically at 38°C for 6 and 24 h with or without CD‐M using hay plus concentrate (1.5:1) as a substrate. The CD‐M was added at different concentrations (0, 1.25, 2.5, 5.0 and 7.5 g/L). The pH of the medium and numbers of protozoa were not affected by the addition of CD‐M. Total volatile fatty acids were increased and ammonia‐N was decreased, molar proportion of acetate was decreased and propionate was increased (P < 0.05) by CD‐M. Methane was inhibited (P < 0.05) by 14–76%. The effect of CD‐M on methane production and ruminal fermentation was further investigated in vivo using four Holstein steers in a cross‐over design. The steers were fed Sudangrass hay and concentrate mixture (1.5:1) with or without CD‐M (2% of feed dry matter) as a supplement. Ruminal proportion of acetate tended to decrease and that of propionate was increased (P < 0.05) 2 h after CD‐M dosing. Total viable counts, cellulolytic, sulfate reducing, acetogenic bacteria and protozoa were unaffected while methanogenic bacteria were decreased (P < 0.05) by CD‐M. The plasma concentration of glucose was increased, whereas that of urea‐N was decreased (P < 0.05). Methane was inhibited (P < 0.05) from 36.4 to 30.1 L/kg dry matter intake by the addition of CD‐M. Apparent digestibilities of dry matter and neutral detergent fiber were not affected while that of crude protein was increased (P < 0.05) in the medicated steers. These data suggested that dietary supplementation of CD‐M decreased methane production and improved nutrient use.     

Effects of cyclodextrin-iodopropane complex on methane production, ruminal fermentation and microbes, digestibility and blood metabolites in steers

The effects when adding cyclodextrin‐iodopropane complex (CD‐IP) to a diet, on ruminal fermentation and microbes, digestibility, blood metabolites and methane production, were evaluated using four Holstein steers in a cross‐over design. The steers were fed Sudangrass hay plus concentrate mixture at a ratio 1.5:1, and CD‐IP (1% of dry matter) was given twice daily by mixing with concentrate mixture. Rumen and blood samples were collected at 0, 2, and 5 h after morning dosing. Ruminal pH and numbers of protozoa were unaffected by CD‐IP treatment. Ruminal molar proportion of acetate was decreased (P < 0.05), and propionate was increased (P < 0.01) at 2 h after CD‐IP dosing. Proportion of butyrate was increased (P < 0.05) and ammonia‐N was decreased (P < 0.05) at 2 and 5 h after CD‐IP dosing. Adding CD‐IP had no effect on the feed intake and digestion of nutrients. Plasma glucose was increased and urea‐N was decreased (P < 0.05) at 2 and 5 h after CD‐IP dosing. Methane production was decreased (P < 0.05) by approximately 18% in the treatment steers. Numbers of methanogenic bacteria were decreased (P < 0.05), while total viable counts, cellulolytic, sulfate reducing and acetogenic bacteria were unaffected. The present results are the first to show that CD‐IP can partially inhibit in vivo ruminal methanogenesis without adverse effects on digestion of nutrients.     

Adherence of Actinobacillus pleuropneumoniae serotype 1 to swine buccal epithelial cells involves fibronectin

The swine pathogen Actinobacillus pleuropneumoniae serotype 1 was investigated for its ability to adhere to swine, rat, and human buccal epithelial cells (BEC). The highest number of bacteria adhered was to swine BEC. This binding ability was affected by heating, extreme pH, treatment with sodium dodecyl sulfate, ethylenediamine tetraacetate, or periodate, and proteolysis, suggesting that cell-surface glycoproteins participate in adherence and that adherence is based mostly on ionic interactions. Mannose and swine fibronectin may play a direct role in this interaction. Convalescent-phase serum from naturally infected pigs inhibited the adhesion. There was a correlation between bacterial pathogenicity as well as host specificity and the capacity for adherence to swine BEC. Adhesion to swine BEC provides a convenient method to study in vitro the adherence of A. pleuropneumoniae and other pathogens of the pig respiratory tract.     

In situ provision of drinking water to grazing dairy cows improves milk production

AIMS: To determine the effect of providing water within the area grazed by dairy cows on milk yield and quality, compared to requiring cows to walk to a distant water trough, on a dairy farm in the Pampa region of Argentina during summer.

METHODS: Holstein dairy cows were allocated to two herds with similar parity, days in milk and milk production. They were grazed in one paddock that was divided in two, with a fixed water trough at one end. Cows were moved twice daily to grazing plots within the paddock. Control cows (n=66) could only access water from the fixed trough, whereas supplemented cows (n=67) also received water from a mobile trough within the grazing plot. Milk production of each cow, and water consumption of the two herds were measured daily over 62 days. Milk composition for each herd was determined weekly from Days 18 to 60 of the study, and grazing behaviour was observed between 08:00 and 16:00 hours on Days 11–15, 19–22 and 39–43.

RESULTS: Over the 62 days of the study, supplemented cows produced 1.39 (SE 0.11) L/cow/day more milk than Control cows (p=0.027). Estimated mean daily water intake was 50.4 (SE 2.1) L/cow/day for supplemented cows and 58.2 (SE 2.7) L/cow/day for Control cows (p=0.004). Percentage total solids in milk was higher for supplemented (12.5 (SE 0.06)%) than Control (12.4 (SE 0.04)%) cows (p=0.047). During the periods of behavioural observation, a higher percentage of cows in the water supplemented than the Control herd were observed in the grazing area (p=0.012).

CONCLUSIONS AND CLINICAL RELEVANCE: This preliminary study demonstrated that provision of water to dairy cows within the grazing plot was beneficial for milk production and composition, and may be associated with longer periods spent within the grazing area, during hot weather in the Pampa region of Argentina.     

Effects of protein-energetic supplementation frequency on growth performance and nutritional characteristics of grazing beef cattle

Two experiments were conducted to evaluate reduced supplementation frequencies for grazing beef cattle in rainy season. In experiment 1, evaluating the nutritional parameters, four rumen-cannulated Nellore bulls (BW = 410 kg) were used. In experiment 2, evaluating animal performance, 48 Nellore bulls (BW = 358 kg) were used. The treatments were as follows: mineral supplement (MS) alone and MS plus protein-energy supplement provided 3×, 5× and 7×/week. Supplementation frequency did not affect (P > 0.05) intake and digestibility. Average daily gain was greater (P < 0.001) to supplementation compared with MS. The supplementation 5×/week resulted in greater weight gain per hectare (9.24) and higher economic returns during the study period (1.64%) compared to other supplementations. Supplementation 5×/week increased animal performance and positively influenced economic returns.     

Interactions between different media and follicle‐stimulating hormone supplementation on in vitro culture of preantral follicles enclosed in ovarian tissue derived from collared peccaries (Pecari tajacu Linneaus, 1758)

The aim was to verify the effect of follicle‐stimulating hormone (FSH) supplementation to α‐MEM+ or TCM199+ media on the in vitro development of ovarian preantral follicles (PFs) derived from collared peccaries. Ovaries (n = 5 pairs) were collected and divided into fragments destined to control group (non‐cultured) or treatments that were cultured for 7 days. The PFs morphology, growth and activation were evaluated by classical histology. The immunohistochemistry markers Ag‐NOR and PCNA were used for nuclear proliferation analysis, and the picrosirius red labelling was used for ovarian extracellular matrix (ECM) evaluation. After 7‐day culture, only the TCM199+ treatment maintained the proportion of intact PFs similar to day 1 (63.2%), but no differences were found among treatments (p > .05). In addition, a significant increase in the growing follicles proportion was verified for all the treatments, indicating follicular activation (p > .05). By the Ag‐NOR analysis, only the TCM199+/FSH maintained the nuclear proliferation similar to the first day (p > .05). The picrosirius red staining revealed that the ECM remained intact in all the treatments (p > .05). We suggest the use of TCM199+ medium supplemented of FSH for the in vitro development of peccaries PFs under 7‐day culturing conditions.     

ACE inhibition in goats under fixed‐time artificial insemination protocol increases the pregnancy rate and twin births

To assess the effect of the angiotensin‐converting enzyme (ACE) inhibition on the efficiency of the fixed‐time artificial insemination (TAI), 69 goats were divided randomly into two groups: enalapril (n = 35) and control (n = 34). In the experiment, all animals underwent the protocol of fixed‐time artificial insemination for 12 days. Enalapril group received enalapril maleate dissolved in saline (Enalapril, Lab Teuto Ltda) subcutaneously at the following doses: 0.2 mg/kg/day in D0‐D2; 0.3 mg/kg/day in D3‐D6 and 0.4 mg/kg/day in D7‐D11. The control group received the corresponding volume of 0.9% saline solution. We performed a single insemination 36 hr after sponge removal using frozen semen from two adult male goats with recognized fertility. The ultrasound pregnancy diagnosis was 30 days after the artificial insemination (AI). There was significant increase in pregnancy rates and twinning as well as a decrease in foetal loss in animals receiving enalapril (p < .01). The use of ACE inhibitors during the TAI protocol was shown to be a promising alternative to increase the efficiency of such reproductive biotechnology.     

The use of ultrasonography in the reproductive evaluation of boars

The objective was to study the use of ultrasound as a complementary test in the breeding soundness evaluation in male pigs and study the pattern of echogenicity of the testicular parenchyma in boars of different racial groups. Twenty‐six adult boars from four different racial groups were used, 10 from the Piau breed (group 1), four from the commercial and finishing group (group 2), six Pietrain breed (group 3) and six from the Duroc breed (group 4). All animals were evaluated for breeding soundness evaluation and the ultrasound examination of the testicles. The groups of animals that were evaluated showed no difference in the main semen parameters that were evaluated, except for the sperm volume, concentration of the ejaculated sperm and the supravital staining; the lowest figures were for the animals from the Piau breed (group 1). In relation to the testicular biometrics, Duroc animals (group 4) had a greater scrotal width compared to the other groups. But when we assessed the intensity of pixels of the testicles, there was a difference between groups. The groups 2 (finishing animals), 3 (Pietrain) and 4 had no difference between themselves. Group 3 had greater pixel intensity in relation to group 1. Of the 26 animals studied, five showed an abnormality during ultrasound evaluation, like hydrocele, hyperechoic mass in the testicular parenchyma, cyst in the head of the epididymis and the presence of fluid in the head and tail of the epididymis. The various animal groups studied did not differ in the principal reproductive parameters evaluated, showing that despite the great variability of reproductive traits between breeds and within the same breed, the breeding soundness evaluation, the more complete it is, is essential for the selection of breeders and the ultrasonography of the reproductive system becomes an important addition in this examination.     

Soil CO2 emission in sugarcane management systems

Sugarcane management systems affect soil attributes such as the carbon cycle. This fact has stimulated the sugar and alcohol industry to refine the sugarcane production systems by replacing the pre-harvest burning (PB) and manual harvest with mechanized harvesting followed by residue deposition. The aim of this study was to evaluate different management systems with respect to C cycling carbon dioxide and soil parameters (chemical, physical and biological) which were determined over the season. Three sugarcane cultivation systems were evaluated at the following periods: (a) PB, (b) 5-year green harvest and (c) 10-year green harvest. The results indicated that CO₂ emission was 36% greater in the 10-year sugarcane green harvest system than in the PB system. The bulk density and macroporosity were the factors that were most affected by the different sugarcane management systems and that significantly influenced soil CO₂ emissions. The principal component analysis showed that soil CO₂ emission was 18% influenced by base saturation (V%) and 14% by pH, especially in the PB area. Additionally, 19% was affected by carbon and macroporosity in the 5- and 10-year green harvest areas, respectively. From our results, it can be concluded that the most CO₂ emissions are in the areas of sugarcane green, this is due to the higher carbon concentration when compared with the area of burning sugarcane. The parameters that most influenced the CO₂ emissions were bulk density, porosity, macroporosity, pH and V%.