Post-thaw Survival and Longevity of Bull Spermatozoa Frozen with an Egg Yolk-based or Two Egg Yolk-free Extenders after an Equilibration Period of 18 h

The aim of the present study was to determine the suitability of using two egg yolk-free commercial extenders, Andromed and Biociphos Plus as compared with the Tris-egg yolk based diluent Biladyl, for the cryopreservation of bull spermatozoa when the freezing protocol involved holding the extended semen at 4 degrees C for 18 h before the freezing. Six ejaculates from each of 10 Holstein bulls were collected by using artificial vagina. The ejaculates were evaluated for volume, sperm concentration and motility, divided in to three equal volumes, and diluted, respectively, with the three extenders as specified above. Extended semen was equilibrated for 18 h at 4 degrees C and frozen in 0.25-ml straws. After thawing, 100-mul aliquots of semen were labelled with SYBR-14, PI and PE-PNA (Phycoerythrin-conjugated Peanut agglutinin) and analysed by flow cytometry at 0, 3, 6 and 9 h after incubation at 37 degrees C. A General Lineal Model procedure for repeated measures was used to determine the effects of extender, bull, replicate and the interaction between them, on sperm viability and acrosomal integrity. Semen samples frozen with Biladyl showed higher (p < 0.001) sperm survival after 0 h (47.9%) and 9 h (30.3%) of incubation than those frozen with Andromed (38.5% and 17.3%, after 0 and 9 h respectively) or Biociphos Plus (34.9% and 21.6%, after 0 and 9 h respectively). The bull and replicate had significant effects (p < 0.001) on both sperm viability and acrosomal integrity, but the interactions between bull and extender and between replicate and extender were not significant. It was concluded that, when holding the semen overnight before freezing, the use of Biladyl results in higher sperm survival and longevity than the use of Andromed or Biociphos Plus.     

Protection of olive planting stocks against parasitism of root-knot nematodes by arbuscular mycorrhizal fungi

The effects were investigated, under controlled conditions, of single and joint inoculation of olive planting stocks cvs Arbequina and Picual with the arbuscular mycorrhizal fungi (AMF) Glomus intraradices , Glomus mosseae or Glomus viscosum , and the root-knot nematodes Meloidogyne incognita and Meloidogyne javanica , on plant performance and nematode infection. Establishment of the fungal symbiosis significantly increased growth of olive plants by 88·9% within a range of 11·9–214·0%, irrespective of olive cultivar, plant age and infection by M. incognita or M. javanica . In plants free from AMF, infection by Meloidogyne spp. significantly reduced the plant main stem diameter by 22·8–38·6%, irrespective of cultivar and plant age. Establishment of AMF in olive plants significantly reduced severity of root galling by 6·3–36·8% as well as reproduction of both Meloidogyne spp. by 11·8–35·7%, indicating a protective effect against parasitism by root-knot nematodes. Infection by the nematodes influenced root colonization by AMF, but the net effect depended on the AMF isolate–olive cultivar combination. It is concluded that prior inoculation of olive plants with AMF may contribute to improving the health status and vigour of cvs Arbequina and Picual planting stocks during nursery propagation.     

Electro-acupuncture and Chinese herbs for treatment of cervical intervertebral disk disease in a dog

A non-ambulatory dog with tetraparesis following a pain episode that had evolved over 2 months was submitted for medical treatment and diagnosed with intervertebral disk disease at C3-C4 and dorsal extradural compression at C1-C2 and C3-C4 using myelography and computed tomography. The dog experienced ambulation recovery after 15 days of treatment with only electroacupuncture and Chinese herbal medicine, with marked improvement occurring after only 10 treatments. Six months of follow-up demonstrated that the dog was stable and had no recurrence of symptoms. Therefore, it was concluded that the combination of electroacupuncture and Chinese herbal medicine was responsible for motor rehabilitation.     

Alternative low doses and routes of administering a prostaglandin F2�� analogue to induce luteolysis in Nelore cows

The present study was conducted in order to verify the efficacy of lower doses and alternative routes of a prostaglandin F2α analogue, luprostiol (PGF), for the induction of luteolysis and the precipitation of estrus in nonlactating Nelore cows (Bos taurus indicus). A conventional dose (15 mg) of PGF was compared to doses lower than the conventional dose, which ranges from 10 to 50%, that were administered intramuscularly (IM), intravulvosubmucosally (IVSM), or in the Bai-hui acupuncture site located within the lumbosacral area. The cows were administered PGF 8 day after estrus in the presence of a corpus luteum, and randomly assigned to the following groups: G1 (positive control), 15 mg, IM (n = 23); G2, 7.5 mg, IM (n = 23); G3, 3.75 mg, IM (n = 24); G4, 7.5 mg, IVSM (n = 25); G5, 3.75 mg, Bai-hui acupoint (n = 24); and G6, 1.5 mg, Bai-hui acupoint (n = 25). The results indicated that 50% of a conventional dose of PGF (7.5 mg) resulted in a complete luteal regression (plasma progesterone <1 ng/ml) at Hour 48, and hastened estrus, regardless of whether or not PGF was administered IM or IVSM. Comparatively, 10 or 25% of the conventional dose, even when administered to the Bai-hui acupoint, resulted in an initial reduction in the concentration of progesterone at Hour 24, followed by an increase observed at Hour 48. In conclusion, 25% of a conventional PGF dose administered via the Bai-hui acupoint proved inadequate to induce a complete luteal regression, whereas 50% of a conventional dose administered IM or IVSM was found to be the minimal dose required to induce effectively a complete luteal regression, and to precipitate the onset of estrus in nonlactating Nelore cows.     

Validation of RT‐PCR Assays for Molecular Characterization of Porcine Teschoviruses and Enteroviruses

Porcine enteroviruses (PEVs) and teschoviruses (PTVs) are described as causative agents of neurological disorders, fertility disorders and dermal lesions of swine. Difficulties in the serological detection of these viruses may lead to a significant underestimation of infections with clinical symptoms. With the recent availability of genome sequence data for all the serotypes, molecular diagnosis is a possibility. The present study describes a new approach to molecular ‘serotyping’ of PTVs and PEV‐B viruses, involving the amplification and sequencing of a genomic fragment of the VP1 coding region. A molecular characterization of Italian entero‐teschovirus isolates was performed using a set of previously published and newly designed polymerase chain reaction primers. A total of 33 porcine isolates and 10 reference strains were analysed. Porcine enterovirus‐B samples were first diagnosed as positive for enterovirus by amplification of the 5′‐non‐translated region. Samples were then typed by amplification and sequencing of a portion of the VP1 coding region. Porcine enterovirus‐A and PTVs were detected by a published assay in the 5′‐NC region that allows them to be differentiated according to the size of amplification product, using the same set of primers. For serotype characterization of PTV, we evaluated four different regions: the N terminus of the capsid protein VP2, the region encoding for RNA‐dependent RNA polymerase, and the capsid VP1 and VP4 regions. The newly designed primers in the VP1 region was proved to be broad in range and suitable for serotype assessment and therefore constitute a useful diagnostic tool for molecular diagnosis of porcine teschovirus/enterovirus strains and for the study of molecular epidemiology and evolution of these viruses.     

Phenotypic characterisation of Phaeoacremonium and Phaeomoniella strains isolated from grapevines: enzyme production and virulence of extra-cellular filtrate on grapevine calluses

Extra-cellular enzyme production of different Phaeoacremonium spp. and Phaemoniella chlamydospora isolates were used to assay the possibility of inter-specific characterisation. Isolates of Phaeoacremonium aleophilum, Phaeoacremonium angustius, P. viticola and Ph. chlamydospora were grown on solid media and the activities of extra-cellular amylases, lipases, proteases, cellulases, xylanases, laccase, polygalacturonase, pectate lyase, lignin peroxidase, manganese peroxidase, urease and chitinase were assayed. Phaeoacremonium species showed activities of a larger number of enzymes and also enzyme activity was frequently higher suggesting that Phaeoacremonium can be more virulent. To assay if the produced extra-cellular enzymes could reflect the virulence capacity of the two genera, calluses of Vitis vinifera L. (cvs. Baga and Maria Gomes) and of a rootstock (R3309) were inoculated with filtrated culture liquid medium of three isolates of Ph. chlamydospora and one of P. angustius. Filtrates from all strains decreased callus growth and membrane integrity, while soluble protein content of calluses decreased with the strains CAP 054 and 1AS. P. angustius (CAP 054) induced the more severe symptoms in all genotypes. Water content decreased together with an increase of osmolality in both cultivars but not in rootstock suggesting that osmorregulatory capacity is more affected in cultivars. Data show that: (1) Phaeoacremonium and Phaeomoniella genera have different patterns of extra- cellular enzymatic production; (2) these fungi produce extra-cellular compound(s) that induce(s) senescence symptoms in plant cells inhibiting callus proliferation; (3) among the strains tested in plant calluses the most virulent isolate (CAP 054) also produced higher amounts of some extra-cellular enzymes; (5) rootstock calluses were less sensitive to inoculation than grapevine calluses.     

A comparison of nitrogen use efficiency definitions in Citrus rootstocks

Nitrogen use efficiency (NUE) definitions, commonly used in literature, are evaluated in response to nitrate availability in four citrus rootstocks, Rough Lemon (Citrus jambhiri Lush) (RL), Sweet Orange (Citrus sinensis (L.) Osbeck) (SwO), Cleopatra Mandarin (Citrus reshni Hort ex Tan.) (CM) and Sour Orange (Citrus aurantium L.) (SO). The application of diverse definitions determine different characterizations in N-efficiency among rootstocks. Nitrogen utilization efficiency (NUtE) and nitrogen efficiency ratio (NER) determine equal level of nitrogen efficiency among all rootstocks. Total nitrogen accumulation (TNA), nitrogen uptake efficiency (NUpE), shoot dry weight (SDW) and total leaf area (TLA) response curves produce the same NUE characterization: SO and SwO were nitrate use efficient and inefficient rootstocks, respectively, while the RL and CM exhibit superior and inferior genetic potential, respectively.     

Effects of deltamethrin on hematological parameters and enzymatic activity in Ancistrus multispinis (Pisces, Teleostei)

Deltamethrin (DM) is a pyrethroid insecticide widely used in Brazilian crops and in pest-control programs because of its low environmental persistence and toxicity. It has been shown to exert a wide range of effects on non-targeted organisms including fishes. The aim of this study to evaluate the effects of deltamethrin through the hematological and biochemical parameters using Ancistrus multispinis as animal model. First, blood of A. multispinis was collected by cardiac puncture in order to evaluate the basal values of the total cell counts, hemoglobin and hematocrit rates. A. multispinis showed low number leukocytes, red blood cells and hemoglobin compared to other species. The intoxication with DM (0.1 or 0.3 mg kg⁻¹) induced leukocytosis and increases the number of erythrocytes and hemoglobin levels 96 h after the injection when compared to vehicle-injected animals. The injection of lipopolysaccharide (LPS) also induced a significant leukocytosis and increased the erythrocytes number 4 h after the injection. The combination of the higher dose of DM with LPS showed an additional effect in the number of leukocytes and erythrocytes. The GST and EROD activities were also evaluated in the gills and liver, respectively, 96 h after intoxication with both doses of DM. The GST activity decreased after DM intoxication in both doses while EROD activity was not significantly different. These results suggest that A. multispinis is a valuable model for ecotoxicological studies. Also, DM can affect the metabolizing system (GST) and immunologic system of A. multispinis, a condition that can be worsened in the presence of inflammation or infection.     

Effects of deficit irrigation strategies on cluster microclimate for improving fruit composition of Moscatel field-grown grapevines

The grapevine plays a very important role in the economic, social and cultural sectors of many regions; however vineyards are often grown in regions under stressful conditions and thus they are vulnerable to climate change. The objective of this research was to investigate the effect of partial root-zone drying (PRD) irrigation on vine water relations, vegetative growth, plant microclimate, berry composition and yield components, compared to conventional deficit irrigation (DI, 50% ETc), full irrigation (FI, 100% of ETc) and non-irrigated vines (NI). The study was undertaken in mature ‘Moscatel’ grapevines (Vitis vinifera L.) grown in Pegões, South of Portugal. Compared to the other irrigated treatments, PRD vines showed a better microclimate at the cluster zone with higher incident photosynthetic photon flux density (PPFD). Within the more open canopies of NI and PRD treatments, berry temperatures were higher than those of denser ones (DI and FI). Compared to the conventional irrigation technique the better microclimate observed in PRD vines was a consequence of a reduction in vine growth, where lower values of leaf layer number, leaf area, canopy wideness, water shoots and shoot weight were observed. In PRD vines we observed a tendency to a development of a deeper root system, while DI and FI showed a more homogeneous root distribution throughout the different soil layers. PRD showed an improvement in berry quality with higher values of flavour precursors, and total phenols concentration without any significant yield reduction compared to DI and FI.     

Plant growth,yield, and fruit quality of ‘Fallglo’ and ‘Sunburst’ mandarins on four rootstocks

Vegetative growth, yield, and fruit quality of ‘Fallglo’ and ‘Sunburst’ mandarins on ‘Rangpur’ lime, ‘Swingle’ citrumelo, ‘Orlando’ tangelo, and ‘Cleopatra’ mandarin were evaluated under subtropical climate of Northern São Paulo State, Brazil, from 2000 through 2006 harvest seasons. ‘Fallglo’ mandarin trees had the highest cumulative yield on ‘Rangpur’ lime, and the smallest on ‘Swingle’ citrumelo and ‘Orlando’ tangelo. Plants of this cultivar had the highest yield efficiency on ‘Rangpur’ lime, and the lowest on ‘Orlando’ tangelo. ‘Sunburst’ mandarin trees began to bear fruits later than ‘Fallglo’ mandarin trees, with no differences in yield induced by the rootstocks. ‘Cleopatra’ mandarin induced the most vigorous growth in ‘Fallglo’ mandarin as compared to plants on ‘Swingle’ citrumelo. On the other hand, the largest trees of ‘Sunburst’ mandarin were registered on ‘Orlando’ tangelo, and the smallest on ‘Rangpur’ lime. ‘Sunburst’ mandarin had higher alternate bearing than ‘Fallglo’ regardless the rootstock. Fruit weight and juice content were not affected by the rootstock. These two mandarin scion cultivars may be considered adequate alternatives to produce good fruit quality for the fresh fruit market. ‘Cleopatra’ mandarin and ‘Rangpur lime are suitable rootstocks for ‘Fallglo’ mandarin, whereas all rootstocks evaluated are adequate for ‘Sunburst’ mandarin.