Isolation and molecular characterisation of Neospora caninum in cattle in New Zealand

AIM: To isolate Neospora caninum from the brains of naturally infected cattle and use molecular techniques to characterise the isolates.

METHODS: Neospora caninum tachyzoites were isolated in Vero cell culture from the brains of a cow and two calves. The isolates were characterised using polymerase chain reaction (PCR) methods, DNA sequencing, an immunofluorescent anti-body test (IFAT), transmission electron microscopy (TEM), and immunohistochemistry (IHC). The brains of the three cattle were subjected to histopathological examination. A pathogenicity study was conducted in 120 BALB/c mice.

RESULTS: Neospora caninum tachyzoites were isolated from all three cases and first observed in vitro between 14 and 17 days post-inoculation. Parasites were sub-cultured and maintained in Vero cell culture for more than 6 months. PCR products were generated for all three isolates, using two different primers. Sequencing of the PCR products and a subsequent BLAST search identified the isolates as N. caninum. In addition, the isolates tested positive using IFAT and IHC, and ultrastructure revealed by TEM was characteristic of N. caninum. Histopathological examination revealed lesions characteristic of N. caninum in 1/3 brains. In the pathogenicity study using BALB/c mice, the mortality rate was 3–7%.

CONCLUSION: This was the first successful isolation of N. caninum in New Zealand confirmed using molecular characterisation tests.     

Nitrogen fluxes during the initial stage of willows and poplars in short‐rotation coppices

Among energy crops, short‐rotation coppices (SRC) are recommended to provide renewable energy. Compared to annual crops, willows and poplars are regarded as plants with low requirements for nutrients, herbicides, pesticides, and soil maintenance. However, only little is known about N‐fertilizer effects on SRC and the few studies are even inconsistent. Therefore, we studied the effects of N on yields of willows and poplars in a field experiment. The effects of N fertilization on nitrate leaching and nitrous oxide emissions from the loamy‐sand soil were also measured. Cuttings of willows (Salix viminalis clone Inger) and poplars (Populus maximovizcii × P. nigra clone max 4) were planted on farmland in 2008. The experiment was arranged in a random block design with three levels of N fertilizer (0, 50, and 75 kg N ha^–1 y^–1). After 2 y, the trees were harvested for the first time. Fertilization did not affect the yields of willows or poplars. However, the application of 75 kg N ha^–1 y^–1 caused an average increase of N leaching in the willow and poplar plots of 25 kg N ha^–1 y^–1 and 40 kg N ha^–1 y^–1, respectively. Emissions of N₂O were increased by a maximum of only 0.2 kg N ha^–1 y^–1. Further, the N fertilizer stimulated the growth of the weed biomass in case of the willow plots by 46% and of the weed N content by 52% (r = 0.53). In conclusion, in the first 2 y, SRC could be produced in a more effective and environmentally friendly manner without mineral fertilizer.     

The fatty acid composition of Nereis diversicolor cultured in an integrated recirculated system: Possible implications for aquaculture

The potential of solid waste originating from a recirculated fish culture system, i.e. faecal material, uneaten food pellets and bacterial biofilms was examined as food source for the marine polychaete Nereis diversicolor. These polychaetes could be a valuable food for fish if they provide essential fatty acids to the fish. Therefore, we analysed the fatty acid profiles from feed and faecal materials, the sediment as well as the cultured organisms — fish and several batches of N. diversicolor — from an integrated recirculating aquaculture system.The major fatty acids (saturated, monounsaturated and polyunsaturated) for all analysed fish feed, fish and faeces samples were C16:0, C18:1 and C22:6 (n − 3), accounting for 48% to 57% of the fatty acids in the samples. The major fatty acids within the sediment were C16:0, C18:1 and C18:3 (n − 3), accounting for 61% of the total fatty acids. The samples of N. diversicolor revealed C16:0, C18:1 and C20:5 (n − 3) as the major fatty acids. Combined, they accounted for 56% of the total fatty acids detected within the worm samples.The results indicate that a recycling or even an upgrade of excreted feed nutrients such as fatty acids, which were otherwise discharged, can be achieved through integrated aquaculture combining fish and worm culture.     

Uranium series dating of human skeletal remains from the del mar and sunnyvale sites, california

Uranium series analyses of human bone samples from the Del Mar and Sunnyvale sites indicate ages of 11,000 and 8,300 years, respectively. The dates are supported by internal concordancy between thorium-230 and protactinium-231 decay systems. These ages are significantly younger than the estimates of 48,000 and 70,000 years based on amino acid racemization, and indicate that the individuals could derive from the population waves that came across the Bering Strait during the last sea-level low.     

The Ethics and Role of AI with Fresh and Frozen Semen in Dogs

The use of artificial insemination (AI) with fresh semen has resulted in many benefits for the management of dog breeding, but there are disadvantages that can sometimes be overlooked. Furthermore, poorer quality semen arising as a result of cryopreservation necessitates uterine insemination, which raises the potential for surgical insemination. A number of significant ethical concerns have been raised by key stakeholders (such as The Kennel Club and the Royal College of Veterinary Surgeons) about AI per se , but particularly about the use of surgical insemination. This paper discusses the technological development of AI and explores a number of the ethical issues raised by its application to dog breeding. An Ethical Matrix method is used to map the potential ethical issues for key interest groups, namely dogs, breeders, owners, veterinarians and wider society. There are national variations in the way in which institutions have evaluated potential ethical impacts, and this is reflected in the different regulatory frameworks governing the use of AI in dogs. In order to facilitate decision-making and reduce some of the ethical risks associated with this technology, the veterinary research community could take several proactive steps including: (i) clarifying clinical decision-making processes, (ii) enhancing informed choice among clients and (iii) increasing the knowledge-base of potential impacts of AI.     

Quantification of the DNA Difference,and Separation of X‐ and Y‐Bearing Sperm in Alpacas (Vicugna pacos)

Sperm sexing is an emerging reproductive technology which has been successfully used to produce offspring of a pre‐determined sex in domestic and wildlife species but has yet to be applied to New World camelids. The aims of the present study were to (i) optimize the Hoescht 33342 (H33342) staining concentration for the flow cytometric separation of X and Y chromosome‐bearing alpaca (Vicugna pacos) sperm nuclei, (ii) separate alpaca sperm nuclei into high purity (>90%) populations bearing the X‐ and Y‐chromosome and (iii) determine the DNA difference between X‐ and Y‐bearing sperm in alpacas. Semen was collected from alpacas and sperm nuclei stained with H33342, incubated and analysed using a high‐speed cell sorter (SX‐MoFlo^®). H33342 staining concentrations of 36, 54, 72 or 90 μm did not affect the proportion of correctly oriented sperm nuclei (43.3 ± 3.9, 46.4 ± 3.7, 44.5 ± 4.0 and 51.1 ± 2.5% respectively) nor the speed of sorting (1381 ± 160, 1386 ± 123, 1371 ± 133 and 1379 ± 127 sperm nuclei/s). Sort reanalysis determined high levels of purity for X‐ and Y‐enriched populations (96.6 ± 0.7% and 96.1 ± 1.1% respectively). The DNA difference, based on fluorescence intensity (determined by the SX‐MoFlo^®), was 3.8 ± 0.06%. These data demonstrate for the first time that alpaca sperm nuclei can be separated into high purity populations and the potential for applying sperm sexing technology to New World camelids.     

Einfluss Kombinierter organischer und mineralischer Düngung auf den Ertrag und Rohproteingehalt von Winterweizen und Wintergerste sowie die Rübenqualität im IOSDV Speyer

Im IOSDV‐Speyer wird die Wechselwirkung der organischen und mineralischen Düngung in der Fruchtfolge Zuckerrüben—Winterweizen—Wintergerste geprüft. Der seit 1984 laufende Versuch wurde nach fünf Rotationen (15 Jahren) ausgewertet. Der mit organischen Düngern ausgebrachte Stickstoff wird ertragswirksam und kann teilweise bei der Düngeplanung berücksichtigt werden. Er wirkt sich insbesondere im Ertragsoptimum positiv auf die Rohproteingehalte im Korn von Weizen und Gerste aus. Vom mit der Rübenblattdüngung ausgebrachten Stickstoff werden ca. 25% bei der Folgekultur Winterweizen überwiegend zur Kornbildung wirksam und können angerechnet werden. Die Stallmist—Nachwirkung ist bei Weizen vorwiegend durch Ertragssteigerung sichtbar und führte zu einer höheren N‐Aufhahme von ca. 15 kg/ha. Bei Wintergerste wirkt die Strohdüngung leicht positiv auf den Ertrag und den Rohproteingehalt. Eine N‐Ausgleichsdüngung zum Stroh ist langfristig nicht erforderlich. Die Stallmist—Nachwirkung zeigt sich auch noch bei Wintergerste und war ertragswirksam wie 25 kg N‐Mineraldünger, sie führte in den niedrigen N‐Stufen über den Mehrertrag zu einer um 5–10 kg/ha höheren N‐Aufhahme. Bei Zuckerrüben verläuft die Ertragskurve des bereinigten Zuckerertrages relativ flach, deshalb kommt der Berücksichtigung der Stickstoffnachlieferung aus der organischen Düngung bei der Düngebedarfsermittlung eine besondere Bedeutung zu.     

Evaluation of colonoscopic allergen provocation as a diagnostic tool in dogs with proven food hypersensitivity reactions

OBJECTIVE: To evaluate the colonoscopic allergen provocation (COLAP) test as a new tool for the diagnosis of IgE-mediated food allergy. METHODS: Oral food challenges as well as COLAP testing were performed in a colony of nine research dogs with proven immediate-type food allergic reactions. In addition, COLAP was performed in five healthy dogs. RESULTS: When compared with the oral challenge test, COLAP accurately determined 18 of 23 (73 per cent) positive oral challenge reactions (73 per cent) in dogs with food allergies and was negative in the healthy dogs. CLINICAL SIGNIFICANCE: The accuracy of this new test may be higher than that for gastric sensitivity testing. Therefore, COLAP holds promise as a new test to confirm the diagnosis of suspect IgE-mediated food allergy in dogs.