Novel insecticide polymer chemistry to reduce the enzymatic digestion of a protein pesticide, trypsin modulating oostatic factor (TMOF)

A limiting factor in the use of proteins as insecticides, especially when the site of action is in the insect hemocoel, is protease degradation in the digestive system and hemolymph and movement across the midgut ventriculus. Trypsin modulating oostatic factor (TMOF) is a per os mosquito peptidic larvacide which moves across the digestive system and binds to receptors on the hemolymph side of the gut where the hormone inhibits protease synthesis and food utilization ultimately causing death. In the current study, the in vitro degradation of TMOF by the digestive enzyme, leucine aminopeptidase, was inhibited by conjugation of TMOF-K with aliphatic polyethylene glycol (PEG) polymers. Structure activity studies demonstrated a correlation between the molecular weight of the PEG polymer and resistance to digestion and show proof of concept that aliphatic-PEG protein polymerization can be used to prevent protease degradation of a protein insecticide.     

The apparent in vivo neuromuscular effects of the δ-endotoxin of Bacillus thuringiensis var. israelensis in mice and insects of four orders

Alkaline-dissolved crystal δ-endotoxin from Bacillus thuringiensis var. israelensis (serovar H 14) was injected into mice and seven species of insects representing the orders Lepidoptera, Orthoptera, Coleoptera, Hemiptera, and Diptera. High in vivo toxicity, at 1 to 5 ppm (μg toxin/g body wet wt), was observed with mice and some insects, including some that are not sensitive to the toxin when administered orally. Neuromuscular effects were observed when the toxin was injected directly into the body cavity of the test animals. Biochemical studies suggested that different protein fragments within the crystal δ-endotoxin may be responsible for the majority of the mosquito larvacidal activity and the neurotoxic symptoms observed in larvae of Trichoplusia ni.     

Bovine paravertebral analgesia: radiographic analysis and suggested method for improvement

The use of paravertebral analgesia in cattle is not always successful. Dissection studies in five cadavers indicated that there was a divergence in the courses of the dorsal and ventral branches of the lumbar nerves which can account for the unreliability of conventional techniques. An alternative more certain method of blocking the lumbar nerves is described.     

Seroprevalence of Toxoplasma gondii in mainland and sub-Antarctic New Zealand sea lion (Phocarctos hookeri) populations

AIMS: To investigate the seroprevalence of antibodies to Toxoplasma gondii in New Zealand sea lions (Phocarctos hookeri), as a potential contributor to reproductive failure.

METHODS: Archived sera were sourced from New Zealand sea lions from two recolonising mainland populations in the Otago Peninsula (n=15) and Stewart Island (n=12), as well as a declining population at Enderby Island (n=28) in the New Zealand sub-Antarctic. Sera were tested for antibodies to T. gondii using a commercially available ELISA (with samples considered positive if the sample to positive ratio was?>30%), and latex agglutination test (LAT; with titres ≥1:32 considered positive). Western blot analysis was used to validate the results of a subset of 14 samples.

RESULTS: Five samples from sea lions in mainland locations were confirmed positive for antibodies to T. gondii. Two adult females exhibited high LAT antibody titres (min 1:2048, max 1:4096) on both occasions when sampled 1 and 2 years apart, respectively. No animals from Enderby Island were seropositive.

CONCLUSIONS: Toxoplasma gondii infection is unlikely to be a major contributor to poor reproductive success in New Zealand sea lions. However, continued surveillance is pertinent to assess subclinical and clinical impacts of the parasite on these threatened populations. The commercial tests evaluated here, with further species-specific threshold refinement could provide a fast, inexpensive and reliable indicator of T. gondii exposure in New Zealand sea lions.     

National Swine brucellosis survey

A national survey was conducted in 1985 to investigate the brucellosis status of the Canadian swine herd. Serum samples were collected from cull sows slaughtered over a forty week period in 1985; 15,707 samples were suitable for brucellosis testing, and 48 (0.31%) gave some degree of reaction on the buffered plate agglutination screening test. All 48 samples were negative on the 2-mercaptoethanol and modified complement fixation test. We therefore conclude that the Canadian swine herd remains free of brucellosis.     

Improving high soy feed formulations for the Florida pompano Trachinotus carolinus through newer soy products and carbohydrase supplementation

Florida pompano perform well on soy‐based, low‐animal protein diets. These diets may be improved through the use of enzymes as a feed supplement or as pretreatment of the soy product prior to inclusion. Novel soy products produced through enzyme treatment or fermentation often have higher protein content and lower levels of indigestible carbohydrates. Two trials were conducted to evaluate the potential of enzyme supplements. The first trial evaluated the effects of carbohydrase supplementation to diets based on two different soybean meals (commodity or selectively bred meal). Pompano fed the selected soybean had significantly improved growth rates and feed efficiency, while enzymatic supplementation had no significant effect on fish performance. However, enzyme supplementation increased phosphorus and energy retention when added to diets based on commodity soybean meal. Carbohydrase supplementation improved dry matter digestibility in commodity soy‐based diets but had no significant effect on selected soybean‐based diets. The second growth trial consisted of an increasing inclusion of an enzymatically treated soybean meal product (NutriVance). Results indicate that NutriVance can replace the totality of commodity soybean without impacting fish performance. Digestibility of the commodity soy, NutriVance, and a fermented soybean meal (Pepsoygen) was also evaluated, with no detectable difference in digestibility values.     

Development of a rapid resistance monitoring bioassay for codling moth larvae

BACKGROUND: The codling moth, Cydia pomonella (L.), is one of the most important pests of apple worldwide. Use of insecticides for management of this insect has been extensive and has resulted in resistance development. There are a number of different bioassay methods to monitor for codling moth resistance; however, many are not applicable to new insecticides and most are time consuming. A novel 16‐well plasticware bioassay plate containing lyophilized diet was developed for rapid resistance monitoring of codling moth. RESULTS: The contact insecticides acetamiprid and azinphosmethyl were significantly more toxic to neonates than to fourth instars. However, there was no significant difference in LC₅₀ values between neonates and fourth instars to the ingestion insecticides chlorantraniliprole, methoxyfenozide, novaluron and spinetoram. Field colonies of codling moth were significantly more resistant to methoxyfenozide than susceptible populations. A diagnostic dose of 20 µg mL^?1 (LC₉₉) was established to monitor for codling moth resistance to methoxyfenozide. CONCLUSIONS: The results presented here demonstrate that a novel and rapid bioassay can be used to monitor for codling moth resistance to methoxyfenozide. The bioassay method is relevant to both ingestion and contact insecticides, but a single diagnostic dose, regardless of larval age, is only relevant to ingestion insecticides. Age‐dependent diagnostic doses are likely necessary for contact insecticides. Copyright © 2011 Society of Chemical Industry     

Molecular characterization, ecology, and epidemiology of a novel Tymovirus in Asclepias viridis from Oklahoma

Native virus-plant interactions require more understanding and their study will provide a basis from which to identify potential sources of emerging destructive viruses in crops. A novel tymovirus sequence was detected in Asclepias viridis (green milkweed), a perennial growing in a natural setting in the Tallgrass Prairie Preserve (TGPP) of Oklahoma. It was abundant within and frequent among A. viridis plants and, to varying extents, within other dicotyledonous and one grass (Panicum virgatum) species obtained from the TGPP. Extracts from A. viridis containing the sequence were infectious to a limited number of species. The virus genome was cloned and determined to be closely related to Kennedya yellow mosaic virus. The persistence of the virus within the Oklahoma A. viridis population was monitored for five successive years. Virus was present in a high percentage of plants within representative areas of the TGPP in all years and was spreading to additional plants. Virus was present in regions adjacent to the TGPP but not in plants sampled from central and south-central Oklahoma. Virus was present in the underground caudex of the plant during the winter, suggesting overwintering in this tissue. The RNA sequence encoding the virus coat protein varied considerably between individual plants (≈3%), likely due to drift rather than selection. An infectious clone was constructed and the virus was named Asclepias asymptomatic virus (AsAV) due to the absence of obvious symptoms on A. viridis.     

Generalized calcinosis cutis associated with probable leptospirosis in a dog

A 6.5-year-old male German Shepherd acutely developed renal and hepatic disease. Serology revealed high concentrations of antibodies against Leptospira copenhageni, and a presumptive diagnosis of leptospirosis was made. The dog was successfully treated with antibiotics and supportive care over a 12-day period. Sixty-two days after the initial presentation, alopecia predominantly involving the dorsum and perineal areas developed. The skin lesions expanded over a 20-day period. Histology revealed generalized calcinosis cutis with follicular atrophy. An injection of 0.01 mg kg-1 dexamethasone suppressed serum cortisol concentrations. No treatment was given and lesions resolved over the following 30 days. This is the third case of generalized calcinosis cutis that has developed in an adult dog after severe systemic disease. Both previous cases developed calcinosis cutis in association with blastomycosis. To the authors' knowledge, this is the first report of generalized calcinosis cutis in an adult dog in association with a presumptive bacterial infection.