The molecular identification and genetic diversity of economically important wireworm species (Coleoptera: Elateridae) in Canada

Click beetle larvae (wireworms) are significant crop pests in Europe and North America. In Canada, there are ~30 economically important species which are morphologically difficult to identify, but for which sequence data are lacking. Accurate knowledge of damage-causing species and the population genetics and phylogeography of elaterids will provide insight into their sustainable management. Here, we use interspecific variation in mitochondrial 16S rRNA as a robust method of identification, consider the intra- and interspecific genetic variation of some important Canadian wireworm pests and assess the genetic structure and isolation by distance for a re-emerging major pest species, Hypnoidus bicolor Eschscholtz. Wireworms were sampled from Canada and the USA, identified as morphospecies, and sequenced at the 16S rRNA region (294–442 bp). Within some species unusually high intraspecific genetic distances between samples suggested the possibility of cryptic wireworm species or misidentifications, though this was <1 % for most species. Phylogenetic analyses gave some indication of the likely identity of these ambiguous samples. There was a significant correlation between genetic and geographic distance and significant genetic structuring within and between H. bicolor populations, which appeared to be composed of two species comprising several haplotypes. These data provide a starting point for determining the distribution of damage-causing species throughout Canada. The inclusion of data from other nuclear and mitochondrial loci, and use of sequence data from known adult samples, would further aid identification and relationships of wireworm species.     

Response of the dusky wireworm, Agriotes obscurus (Coleoptera: Elateridae), to residual levels of bifenthrin in field soil

To determine if bifenthrin residues elicit morbidity and surfacing behavior in wireworms, larvae of the dusky wireworm, Agriotes obscurus (Coleoptera: Elateridae) were placed in field soil treated with the pyrethroid insecticide bifenthrin ~1 year previous. Morbidity was immediate and lasted as long as wireworms remained in the soil, disappearing quickly after transfer to clean soil. In 2009, field soil treated 336 days previous with bifenthrin at 340 g AI/ha elicited morbidity symptoms similar to that elicited by soil freshly amended with bifenthrin at 100 g AI, and analysis of the field soil confirmed residual levels of bifenthrin exceeding 100 g AI/ha. In 2010, wireworms placed in field soil treated 343 days previous with bifenthrin at 100, 200, and 300 g AI/ha responded as in 2009, with the degree of morbidity increasing with the rate of insecticide, and with wireworms in a non-feeding state more affected than those in a feeding state at each rate. In both 2009 and 2010, moribund wireworms moved to the soil surface within 1 day of placement in the soil containing residual bifenthrin and remained there until reburied, after which they often resurfaced. To confirm that the bifenthrin residues elicited repellency, wireworms were placed in soil window bioassays containing field soil with residual bifenthrin. Wireworms behaved markedly different upon contacting soil containing the residues than when exposed to untreated soil, both in the presence and absence of an attractant, but were less likely to avoid soil containing residual bifenthrin when attracted by wheat seedlings placed inside it.     

Influence of ante‐ and peri‐mortem factors on biochemical and physical characteristics of turkey breast muscle

Summary

The course of post‐mortem breakdown of glycogen and ATP in turkey pectoralis major muscle was markedly influenced by several ante‐ and peri‐mortem variables. Application of a proper stunning procedure was highly effective in preventing peri‐ and post‐mortem muscle stress reactions.

The physiological level of glycogen and ATP was not significantly affected by road transportation covering 260 km. Birds which rested for 24 hrs following transportation had lower glycogen and ATP levels at the moment of slaughter than non‐rested birds.

According to the changes in the rate and extent of post‐mortem biochemical reactions, several meat characteristics such as water‐holding capacity, colour, and tenderness were significantly changed.

Furthermore, the results also indicate that turkey breast muscle is susceptible to a PSE‐like condition as described in pork.     

Streptococcus suis type 2 infections in pigs in the Netherlands (part two)

Summary

Since 1983 some pig breeding and fattening farms in the Netherlands have been faced with a considerable mortality in pigs due to Streptococcus suis type 2 infections. The most predominant clinical feature of S. suis type 2 infection is meningitis, although sudden deaths often occur. It was noted that some affected farms had imported breeding stock from the United Kingdom.

Tonsils of slaughter pigs were collected from herds with and without a history of S. suis type 2 infections. Bacteriological examination was done by using an elective‐selective medium. No significant difference was found in carrier rates of S. suis type 2 between clinically healthy and affected herds (38% vs. 45%).

A cohort study was carried out by regular bacteriological examination of tonsil biopsies on a farm with a high incidence of streptococcal meningitis. Twenty‐seven percent of the pigs were carriers of S. suis type 2 at nine weeks of age. Possible methods for disease control are discussed.     

Destructive cholangiolitis in seven dogs

Summary

In seven dogs presented with clinical signs and laboratory data suggestive of extrahepatic cholestasis, destructive cholangiolitis was diagnosed. The diagnosis was based on the absence of extrahepatic cholestasis at laparoscopy, laparotomy and/or post‐mortem examination, and the presence of specific liver lesions i.e. loss of bile ducts in the smaller portal areas. The disease is compared with drug‐induced (chlorpromazine) cholestasis in man. In two dogs clinical signs were preceded by longstanding respectively repeated sulphonamide medication.     

Virulence of five live vaccines against Avian Infectious Laryngotracheitis and their immunogenicity and spread after eyedrop or spray application

Summary

Five live virus vaccines against avian infectious laryngotracheitis were studied with regard to safety, immunogenicity and route of administration. Significant differences in virulence between the vaccine strains were found. Reduced virulence was accompanied by a reduction of immunogenicity and capacity to spread.

After eyedrop application, a low virulent vaccine induced 90‐ 100% flock immunity for the first 10 weeks after vaccination (PV), followed by a slow decline to 50% at 31 weeks PV, whereas flock immunity induced with the more virulent types remained at about 90% till the end of the experiments (24 and 48 weeks PV).

Aerosol vaccination induced 70 ‐ 100% flock immunity but vaccine reactions were severe. Application of vaccine in a coarse spray did not result in adverse vaccine reactions but induced a maximal protection rate of only 50%.

Microneutralisation titres provided a useful indicator of immunity from the onset of immunity until immunity started to decline. A vaccine virus carrier state was demonstrated by means of sentinel birds.     

Bilirubin metabolism in canine hepatobiliary and haemolytic disease

Summary

The kinetics of unconjugated ₃H‐bilirubin are described in 25 healthy dogs and 35 dogs with spontaneous hepatobiliary or haemolytic disease, using a two‐compartment model. The bilirubin production rates from erythrocyte degradation (P_E), ineffective erythropoiesis (P_I) and catabolism of hepatic haemoproteins (P_I), were derived from the incorporation of ¹⁴C‐glycine into haemoglobin and stercobilin. These combined measurements permitted an integral survey of bilirubin metabolism in health and disease. The concentration of unconjugated bilirubin in plasma and its of total bilirubin levels were similar in hepatic and haemolytic disorders.

This was explained by the highly increased bilirubin production rates in both types of disease. In addition, the hepatic bilirubin clearance was severely impaired in fulminant hepatitis and in cirrhosis, and moderately decreased in the other hepatobiliary diseases and in primary haemolysis. The erythrocyte lifespan was reduced in all animals but one. In addition to haemolysis, the contribution of P_I and P_L was variable, and in two dogs P_L was the principle source of highly increased bilirubin production rates. These data indicate that the concentration of unconjugated bilirubin in plasma or its fraction of total pigments is unreliable in the discrimination of canine hepatobiliary disease from haemolytic disorders.     

Effect of glucocorticoids on cows suspected of subclinical infection with M. paratuberculosis

Summary

Glucocorticoids were administered to 10 heifers suspected of subclinical infection with Mycobacterium paratuberculosis. Three animals remained untreated.

M. paratuberculosis was isolated from the internal organs of 2 animals after this treatment but not from any of the control group. Delayed type hypersensitivity and lymphocyte reactivity towards Johnin and purified protein derivates of M. avium and M. bovis were depressed. A sharp increase in total leucocyte count, due loan increase in neutrophil numbers, occurred. In the three untreated animals these parameters did not change during the experiment.

A decrease of specific immunological reactivity towards M. paratuberculosis occurred, but not to such an extent that clinical disease developed.