First Established Pregnancies in Mediterranean Italian Buffaloes (Bubalus bubalis) Following Deposition of Sexed Spermatozoa near the Utero Tubal Junction

At the time of AI following Ovsynch protocol, a total of 51 buffaloes were randomly divided in a first group (n = 30) subjected to conventional AI into the uterine body with 20 million non-sex sorted frozen-thawed spermatozoa, while a second group (n = 21) was inseminated near the utero-tubal junction (UTJ) ipsilateral to the ovary carrying the preovulatory follicle with 2.5 million live (4 million total) sex-sorted frozen-thawed spermatozoa. The semen used for flowcytometric sorting was collected and processed on a farm in Italy, and then shipped to a laboratory in Germany. Eleven buffaloes were inseminated with X-chromosome bearing spermatozoa and 10 with Y-chromosome bearing spermatozoa. Conception rates after conventional and UTJ inseminations were 43.3% (n = 13) and 42.8% (n = 9) respectively (p = 0.97). Eight of the nine foetuses obtained after insemination with sexed spermatozoa corresponded to the sex as predicted by the cell sorting procedure (five male and four female foetuses by ultrasound vs six male and three female foetuses by cell sorting). In conclusion, for the first time buffalo semen has been successfully subjected to procedures for flowcytometric sperm sorting and freezing. Low doses of sexed spermatozoa have been deposited near the UTJ giving conception rates similar to those of conventional AI with full dose.     

Impact of foot-and-mouth disease on mastitis and culling on a large-scale dairy farm in Kenya

Foot and mouth disease (FMD) is a highly transmissible viral infection of cloven hooved animals associated with severe economic losses when introduced into FMD-free countries. Information on the impact of the disease in FMDV-endemic countries is poorly characterised yet essential for the prioritisation of scarce resources for disease control programmes. A FMD (virus serotype SAT2) outbreak on a large-scale dairy farm in Nakuru County, Kenya provided an opportunity to evaluate the impact of FMD on clinical mastitis and culling rate. A cohort approach followed animals over a 12-month period after the commencement of the outbreak. For culling, all animals were included; for mastitis, those over 18 months of age. FMD was recorded in 400/644 cattle over a 29-day period. During the follow-up period 76 animals were culled or died whilst in the over 18 month old cohort 63 developed clinical mastitis. Hazard ratios (HR) were generated using Cox regression accounting for non-proportional hazards by inclusion of time-varying effects. Univariable analysis showed FMD cases were culled sooner but there was no effect on clinical mastitis. After adjusting for possible confounders and inclusion of time-varying effects there was weak evidence to support an effect of FMD on culling (HR = 1.7, 95% confidence intervals [CI] 0.88-3.1, P = 0.12). For mastitis, there was stronger evidence of an increased rate in the first month after the onset of the outbreak (HR = 2.9, 95%CI 0.97-8.9, P = 0.057).

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-015-0173-4) contains supplementary material, which is available to authorized users.     

Screening techniques and sources of resistance to abiotic stresses in cool-season food legumes

Summary The adaptability and productivity of cool-season food legumes (chickpea, faba bean, lentil, pea) are limited by major abiotic stresses including drought, heat, frost, chilling, waterlogging, salinity and mineral toxicities. The severity of these stresses is unpredictable in field experiments, so field trials are increasingly supplemented with controlled-environment testing and physiological screening. For drought testing, irrigation is used in dry fields and rain-out shelters in damp ones. Carbon isotope discrimination (Δ¹³C) is a well-established screen for drought tolerance in C3 cereal crops which is now being validated for use in grain legumes, but it is relatively expensive per sample and more economical methods include stomatal conductance and canopy temperature. Chickpea lines ICC4958 and FLIP87-59C and faba bean line ILB938 have demonstrated good drought tolerance parameters in different experiments. For frost tolerance, an efficient controlled-environment procedure involves exposing hardened pot-grown plants to sub-zero temperatures. Faba beans Cote d’Or and BPL4628 as well as lentil ILL5865 have demonstrated good freezing tolerance in such tests. Chilling-tolerance tests are more commonly conducted in the field and lentil line ILL1878 as well as derivatives of interspecific crosses between chickpea and its wild relatives have repeatedly shown good results. The timing of chilling is particularly important as temperatures which are not lethal to the plant can greatly disrupt fertilization of flowers. Salinity response can be determined using hydroponic methods with a sand or gravel substrate and rapid, efficient scoring is based on leaf symptoms. Many lines of chickpea, faba bean and lentil have shown good salinity tolerance in a single article but none has become a benchmark. Waterlogging tolerance can be evaluated using paired hydroponic systems, one oxygenated and the other de-oxygenated. The development of lysigenous cavities or aerenchyma in roots, common in warm-season legumes, is reported in pea and lentil but is not well established in chickpea or faba bean. Many stresses are associated with oxidative damage leading to changes in chlorophyll fluorescence, membrane stability and peroxidase levels. An additional factor relevant to the legumes is the response of the symbiotic nitrogen-fixing bacteria to the stress.     

Pollination, Fertilization and Seed Development in Inbred Lines and F1 Hybrids of Spring Faba Beans (Vicia faba L.)

When the flowers are not disturbed, the yield of F₁ hybrids of faba beans normally exceeds that in their inbred parents. The basis of this “heterotic autofertility” was investigated by examining flowers for pollination and fertilization throughout the flowering season and assessing the distribution of seeds and pods on mature plants. Six inbred lines and seven of their F₁ hybrids were grown without protection from bee visitation, and their autofertility was estimated’ by comparing their fertilization with that of cytoplasmic male-sterile line. The first direct evidence was obtained that heterotic autofertility acts by increasing; the proportion of flowers which were pollinated and fertilized. The autofertility was not perfect, especially in a closed-flower hybrid, as many ovules remained unfertilized. Several factors contributed to the number of seeds per pod in F₁ hybrids, being superior to parental means: (1) more ovules per flower, (2) greater fertilization of the basal ovules, (3) reduced abortion of the apical ovules and (4) greater overall maturation ovules.     

Assessment of Nitrogen Leaching at ICP-Waters Sites (Europe and North America)

The International Cooperative Programme on Assessment and Monitoring of Acidification of Rivers and Lakes (ICP-Waters) is designed to assess the degree and geographical extent of acidification of surface waters. During the last 20 years, international emission reduction measures in Europe and North America have resulted in a decrease in atmospheric S-deposition of up to 50%, while N-deposition has stayed almost constant. Empirical relationships between N-deposition and stages of nitrogen saturation are used to assess the importance of nitrogen leaching at the ICP-Waters sites in Europe and North America. There is a clear coherence between high N saturation stages and high N deposition loads and more than 50% of the analysed ICP Waters sites show a high degree of nitrogen saturation, defined as stages 2 or 3 in Stoddard's classification system. Significant leaching of inorganic N also occurs at most sites with deposition greater than 10 kg/ha/yr.     

Implication of Lake Water Residence Time on the Classification of Norwegian Surface Water Sites into Progressive Stages of Nitrogen Saturation

Seasonal behaviour of NO₃ ^- in surfacewater is often used as an indicator on a catchment's ability toretain N from atmospheric deposition. In this paper, weclassify 12 pristine sites (five streams and seven lakes) insouthernmost Norway according to the N saturation stageconcept. All examined sites were acid-sensitive and had annualmean NO₃ ^- concentrations in the range 6–36 μeqL^-1. At lake sites with relatively long water residencetimes, hydrology will have a damping effect on the seasonalNO₃ ^- oscillations in the outlet streams. Under givenrunoff conditions (1.0–2.2 m yr^-1), the seasonalNO₃ ^- pattern was clearly affected when lake:catchmentarea ratios exceeded 0.15 and the total residence time of lakesin the catchments exceeded 0.3 yr. In such cases, annualmean NO₃ ^- concentration rather than seasonalvariations may be the better indicator of N saturation. Toaccount for this we propose a set of supplementary criteria forsuch lake systems, defining limit values for annual meanNO₃ ^- concentrations within each stage class. Beforebeing applied on a broader scale, however, we recommend anexamination of additional lake-dominated catchmentsrepresenting a larger gradient in ecosystem types, N depositionlevels and hydrologic regimes. When applying the supplementarycriteria on the Norwegian sites, two were classified as stage 0(no saturation), five as stage 1 (early stage of saturation)and five as stage 2 (saturated – moderate N loss). No siteswere showing symptoms of stage 3, which characteriseswatersheds that are net sources of N, rather than sinks. Theresults indicate a great variability in N retention capacity inthe study region, despite the fact that many of the catchmentsexperience relatively uniform N deposition amounts and climaticconditions. This suggests that much of this variability must bedue to specific catchment characteristics as e.g. soil type,soil depth, and vegetation cover.     

Computed tomographic assessment of vascular invasion and resectability of mediastinal masses in dogs and a cat

AIMS: To assess the sensitivity of non-angiographic contrast-enhanced computed tomography (CT) to determine the presence of vascular invasion of cranial mediastinal masses in dogs and a cat, and to evaluate the association between vascular invasion and peri-operative mortality.

METHODS: A retrospective study was conducted on 25 dogs and one cat. CT scans were completed with slices ranging from 2 to 10 mm. CT images were evaluated by a board-certifi ed radiologist blinded to previous diagnoses and surgical fi ndings. Each CT study was evaluated for vascular invasion, defi ned as disruption of the vessel wall and extension of the mass into the vessel lumen. Data retrieved from the surgery reports included surgical approach, whether vascular invasion was present, the surgeon's decision on operability, and post-operative complications.

RESULTS: Computed tomographic evaluation revealed 25/26 masses had no evidence of vascular invasion. During surgical exploration, 10/26 masses were found to invade major regional vasculature; the cranial vena cava (CVC) was the vessel most commonly invaded (7/10 animals), and 4/7 (57%) patients with invasion of the CVC were euthanised or died in the perioperative period, from surgical or disease-related problems, which was signifi cantly higher than patients without vascular invasion (p=0.045).

CONCLUSIONS: Non-angiographic contrast-enhanced CT was signifi cantly less sensitive for detecting vascular invasion of cranial mediastinal masses when compared with surgical evaluation. If the CVC was invaded by a tumour there was a signifi cant risk of death peri-operatively when compared with non-invasive cases.

CLINICAL RELEVANCE: Due to the signifi cantly higher mortality risk associated with invasion of the CVC, a more sensitive method than CT should be investigated to determine vascular invasion of mediastinal masses pre-operatively.     

Macrocyclic lactone toxicity due to abamectin in farm dogs without the ABCB1 gene mutation

Abstract

CASE HISTORY: A 5-year-old entire female Huntaway from a sheep and beef farm was one of four dogs that developed clinical signs including hypersalivation, depression, blindness and ataxia after the death of another dog. A 4-year-old female Huntaway farm dog from a second farm was observed to be sitting down more often than usual on the day after being fed part of a calf carcass that had been treated with an abamectin pour-on.

CLINICAL FINDINGS: The first dog was ataxic and depressed but did respond to sound. The second dog presented with an acute onset of blindness, mydriasis, absence of a menace response, hypersalivation, gait abnormalities (e.g. high stepping gait and ataxia), and depression. Other presenting signs included muscle tremors, dehydration and difficulty eating. No abnormalities were detected from routine haematology and biochemistry. Analysis of samples of plasma from both dogs revealed concentrations of abamectin of 0.149 mg/L and 0.260 mg/L for the first and second dogs, respectively. Buccal swabs taken from the first dog for DNA testing for the ABCB1 gene mutation, gave a negative result.

DIAGNOSIS: In addition to the presenting signs which suggested a toxicosis, both dogs had measurable concentrations of abamectin in plasma confirming exposure.

CLINICAL RELEVANCE: Farm dogs exposed to concentrated pour-on products containing abamectin have been poisoned and recover or die. The product labels do not carry any warnings as to the risk of poisoning to dogs. This paper discusses two incidents affecting six farm dogs, but the authors are aware of more toxicoses in farm dogs exposed to abamectin.