Improving the persistence of a formulation of the avian repellent cinnamamide, for the protection of autumn-sown oilseed rape

In a previous pilot field trial on autumn-sown oilseed rape (Brassica napus L), a suspension concentrate formulation of the avian repellent cinnamamide, using Acronal 4D (50 g litre(-1)) as the sticker, was shown to reduce damage by woodpigeons (Columba palumbus L). However, the persistence of cinnamamide on rape leaves was poor. This paper describes the development of a weather-resistant formulation by the inclusion of a more effective sticker. In vitro studies were used to investigate wash-off of cinnamamide, which was identified as being the major mechanism of loss in the field. Rainfastness of cinnamamide was assessed by washing deposits on glass slides and rape leaves using a 'rain-washing machine'. A number of stickers were evaluated and the effect of sticker concentration, ageing of deposits, freezing and intermittent washing on rain-fastness was investigated. None of the candidate stickers showed increased rainfastness compared to Acronal 4D. However, two of the stickers, Crovol T40 G and Emoleo L1, when combined with Acronal 4D in small amounts, did reduce wash-off compared with Acronal 4D alone. Freezing led to an increase in wash-off of cinnamamide, but inclusion of Emoleo L1 with Acronal 4D reduced this effect. Increasing the concentration of the sticker was also shown to improve the rainfastness; doubling the concentration of Acronal from 50 to 100 g litre(-1) halved the wash-off of cinnamamide.     

Oral doxycycline, niacinamide and prednisolone used to treat bilateral nodular granulomatous conjunctivitis of the third eyelid in an Australian Kelpie dog

A 5-year-old, female neutered, Australian Kelpie presented with a 2-month history of dramatic bilateral erythematous thickening of the third eyelids. Ophthalmic examination demonstrated raised, pink to red, irregular thickening of the entire palpebral surface of both third eyelids. There were no other ocular abnormalities. A surgical biopsy was taken from each third eyelid. Histopathologic examination revealed sheets of macrophages, plasma cells, lymphocytes, and occasional fibroblasts and neutrophils infiltrating the third eyelid stroma. A diagnosis of chronic granulomatous conjunctivitis was made. Grossly and histopathologically this case closely resembles previously described cases of nodular granulomatous conjunctivitis involving the third eyelids of Collie dogs. This report describes an unusual case of nodular granulomatous conjunctivitis isolated to the third eyelids in an Australian Kelpie. Resolution of the condition was achieved with a combination of oral doxycycline, niacinamide and prednisolone.     

Internal parasites of possums (Trichosurus vulpecula) from Kawau Island, Chatham Island and Stewart Island

As part of a search for pathogens that might be useful agents for biological control of possums, the three largest offshore islands of New Zealand that still have possums were surveyed to determine the pathogens present in these isolated populations. Brushtail possums from Kawau Island (n = 158), Chatham Island (n = 214) and Stewart Island (n = 194) were examined for internal parasites. Possums from Kawau Island were infected with Eimeria spp. (16.7%), Bertiella trichosuri (5.2%) and Purustrongyloides trichosuri (15.5%). No Paraustrostrongylus trichosuri or Trichostrongylus spp. were found. Possums from Chatham Island were infected with Eimeria spp. (10.9%). Bertiella trichosuri (3.6%), T colubriformis (6.6%), T retortaeformis (1%) and T. vitrinus (0.5%). No Parastrongyloides or Paraustrostrongylus spp. were found. Possums from Stewart Island were infected only with Eimeria spp. (4.6%). Because of their paucity of some parasites, the opportunity exists to use these offshore islands to study the introduction and spread of a parasite into a possum population, and what technology would be required to bring it to hyperendemicity.     

Foraging and denning patterns of brushtail possums, and their possible relationship to contact with cattle and the transmission of bovine tuberculosis

Radio-tracking and direct observation were used over 18 months in 1990-92 to investigate both the use of sleeping dens and foraging activity by possums (Trichosurus vulpecula) on a 21 ha site in the Wairarapa used for a longitudinal study of bovine tuberculosis. Males had larger home ranges than females, and both sexes had larger activity areas during the autumn mating season than at other times of the year. Possums typically foraged in only a small area of their home ranges (termed an activity area) on any one night, and the areas used by individuals were commonly very similar over a series of nights. Activity areas overlapped extensively among possums. Possums used a limited number of dens, commonly in a small and in most cases a circumscribed part of their home range. No simultaneous den-sharing was found, with the exception of mother-joey pairs. The mortality of juveniles after independence was 36%. Only two of 25 juveniles under surveillance to detect dispersal dispersed more than 500 m off the study site, and both subsequently died. Grazing patterns of cattle meant that almost all accessible areas of the paddock were covered by at least some grazing cattle, and so all activity areas of possums within the paddock were covered by areas where cattle foraged. However, possums avoided contact with cattle, and when some cattle were excluded from access to the part of the paddock principally used by both tuberculous and healthy possums for denning, transmission of Mycobacterium bovis from possums to these cattle ceased, although there was subsequent transmission to deer. Cattle which grazed the area used principally for possum denning continued to become infected, and these denning areas appeared to be of importance in the transmission of tuberculosis.     

Sampling rare and elusive populations

The sampling of rare and elusive populations is difficult because the costs of locating such populations are substantial and can exceed actual interviewing costs. There are efficient probability methods that have been developed recently that reduce these costs. If the special populations are geographically clustered, efficient sampling involves the rapid location of segments in which no members of the special population are located with the use of Census data, telephone screening, or incomplete lists. Populations that are not geographicaily clustered can be located by network sampling and use of large previously gathered samples. Characteristics of mobile populations such as the homeless can be estimated by capture-recapture methods.     

Vesicles Cytoplasmic Injection: An Efficient Technique to Produce Porcine Transgene‐Expressing Embryos

The use of vesicles co‐incubated with plasmids showed to improve the efficiency of cytoplasmic injection of transgenes in cattle. Here, this technique was tested as a simplified alternative for transgenes delivery in porcine zygotes. To this aim, cytoplasmic injection of the plasmid alone was compared to the injection with plasmids co‐incubated with vesicles both in diploid parthenogenic and IVF zygotes. The plasmid pcx‐egfp was injected circular (CP) at 3, 30 and 300 ng/μl and linear (LP) at 30 ng/μl. The experimental groups using parthenogenetic zygotes were as follows: CP naked at 3 ng/μl (N = 105), 30 ng/μl (N = 95) and 300 ng/μl (N = 65); Sham (N = 105); control not injected (N = 223); LP naked at 30 ng/μl (N = 78); LP vesicles (N = 115) and Sham vesicles (N = 59). For IVF zygotes: LP naked (N = 44) LP vesicles (N = 94), Sham (N = 59) and control (N = 79). Cleavage, blastocyst and GFP+ rates were analysed by Fisher's test (p < 0.05). The parthenogenic CP naked group showed lower cleavage respect to control (p < 0.05). The highest concentration of plasmids to allow development to blastocyst stage was 30 ng/μl. There were no differences in DNA fragmentation between groups. The parthenogenic LP naked group resulted in high GFP rates (46%) and also allowed the production of GFP blastocysts (33%). The cytoplasmic injection with LP vesicles into parthenogenic zygotes allowed 100% GFP blastocysts. Injected IVF showed higher cleavage rates than control (p < 0.05). In IVF zygotes, only the use of vesicles produced GFP blastocysts. The use of vesicles co‐incubated with plasmids improves the transgene expression efficiency for cytoplasmic injection in porcine zygotes and constitutes a simple technique for easy delivery of plasmids.     

An improved method for measuring soil N2O fluxes using a quantum cascade laser with a dynamic chamber

A dynamic chamber method was developed to measure fluxes of N₂O from soils with greater accuracy than previously possible, through the use of a quantum cascade laser (QCL). The dynamic method was compared with the conventional static chamber method, where samples are analysed subsequently on a gas chromatograph. Results suggest that the dynamic method is capable of measuring soil N₂O fluxes with an uncertainty of typically less than 1–2 µg N₂O‐N m^?2 hour^?1 (0.24–0.48 g N₂O‐N ha^?1 day^?1), much less than the conventional static chamber method, because of the greater precision and temporal resolution of the QCL. The continuous record of N₂O and CO₂ concentration at 1 Hz during chamber closure provides an insight into the effects that enclosure time and the use of different regression methods may introduce when employed with static chamber systems similar in design. Results suggest that long enclosure times can contribute significantly to uncertainty in chamber flux measurements. Non‐linear models are less influenced by effects of long enclosure time, but even these do not always adequately describe the observed concentrations when enclosure time exceeds 10 minutes, especially with large fluxes.     

Evaluation of Cheetah and Leopard Spermatozoa Developmental Capability after Interspecific ICSI with Domestic Cat Oocytes

The ICSI procedure is potentially of great value for felids, and it has not been extensively studied in these species. The objectives of this work were to determine the best conditions for ICSI in the domestic cat (DC) to generate interspecific embryos by injecting cheetah (Ch) and leopard (Leo) spermatozoa. Firstly, DC oocytes were matured with insulin–transferrin–selenium (ITS) or without it (MM) and cultured using atmospheric (21%) or low (5%) oxygen tension after ICSI. The group ITS‐5%O₂ showed the highest blastocyst rate (p < 0.05), 20.9% vs 8.7%, 7% and 6.5%, for MM‐21%O₂, MM‐5%O₂ and ITS‐21%O₂, respectively. The best conditions were used to generate the interspecific embryos, together with ionomycin activation (Io) after ICSI. Interspecific embryos resulted in high rates of blastocysts that were not positively affected by Io activation: 32.6% vs 21% for Ch and Ch‐Io, 9.8% vs 21% for Leo and Leo‐Io, and 20% vs 17.4% for DC and DC‐Io. We also evaluated DNA‐fragmented nuclei of experiment 1 and 2 blastocysts, using TUNEL assay. The fragmented nucleus proportion was higher in the ITS‐5%O₂ group, 67.6%. Surprisingly, interspecific blastocysts showed the lowest fragmented nucleus proportion: 27% and 29.9% for Ch and Leo, respectively. We concluded that ITS and 5%O₂ improve blastocyst formation in DC, although with a concomitant increase in DNA fragmentation. Most importantly, cheetah and leopard spermatozoa were able to generate blastocysts without artificial activation, which suggests that developmental capacity of wild felid spermatozoa can be evaluated by interspecific ICSI. This technique should be used to assist wild felid reproduction.