Eggert Gunnarsson

Lack of evidence for vertical transmission of Campylobacter spp. in chickens.

ABSTRACT Campylobacter jejuni is a major cause of bacterial food-borne infection in the industrial world. There is evidence that C. jejuni is present in eggs and hatchery fluff, opening the possibility for vertical transmission from hens to progeny. Poultry operations in Iceland provide an excellent opportunity to study this possibility, since breeding flocks are established solely from eggs imported from grandparent flocks in Sweden. This leaves limited opportunity for grandparents and their progeny to share isolates through horizontal transmission. While Campylobacter was not detected in all grandparent flocks, 13 of the 16 egg import lots consisted of eggs gathered from one or more Campylobacter-positive grandparent flocks. No evidence of Campylobacter was found by PCR in any of the 10 relevant quarantine hatchery fluff samples examined, and no Campylobacter was isolated from the parent birds through 8 weeks, while they were still in quarantine rearing facilities. After the birds were moved to less biosecure rearing facilities, Campylobacter was isolated, and 29 alleles were observed among the 224 isolates studied. While three alleles were found in both Sweden and Iceland, in no case was the same allele found both in a particular grandparent flock and in its progeny. We could find no evidence for vertical transmission of Campylobacter to the approximately 60,000 progeny parent breeders that were hatched from eggs coming from Campylobacter-positive grandparent flocks. If vertical transmission is occurring, it is not a significant source for the contamination of chicken flocks with Campylobacter spp.

Paratuberculosis in Iceland : epidemiology and control measures, past and present

Paratuberculosis as well as the slow virus infections maedi/visna and jaagsiekte came to Iceland in 1933 when 20 sheep of the Karakul breed were imported from Halle, Germany. At least five of these sheep were subclinical carriers of paratuberculosis. Within 16 years paratuberculosis together with the other Karakul diseases (maedi/visna and jaagsiekte) almost ruined sheep farming, the main agricultural industry in Iceland. The first clinical case of paratuberculosis in sheep was confirmed in 1938, and in cattle in 1944. The first cattle cases of paratuberculosis appeared on farms where the disease had been prevalent in sheep for years. The virulence in cattle appeared to be considerably lower than in sheep. Extensive measures were used to control the spread of paratuberculosis in sheep. Hundreds of kilometres of fences were put up and used together with natural geographic borders to restrict the movement of sheep from infected areas. Serological and other immunological tests were also used to detect and dispose of infected individuals. These measures proved inadequate and the disease could not be eradicated. Culling and restocking of uninfected sheep in endemic areas eradicated maedi/visna and jaagsiekte but not paratuberculosis. Experiments showed that vaccination against paratuberculosis could reduce mortality in sheep by 94%. Vaccination of sheep in endemic areas has been compulsory in Iceland since 1966 and as a result losses have been reduced considerably. Today, serology is used to detect and control infection in cattle herds. Furthermore, serology is used to control vaccination of sheep and screen for infection in non-endemic areas. The complement fixation (CF) test for paratuberculosis has been used until now, but recently we have started comparing the CF test with the CSL absorbed ELISA test.

Broiler Campylobacter Contamination and Human Campylobacteriosis in Iceland

ABSTRACT To examine whether there is a relationship between the degree of Campylobacter contamination observed in product lots of retail Icelandic broiler chicken carcasses and the incidence of human disease, 1,617 isolates from 327 individual product lots were genetically matched (using the flaA short variable region [SVR[) to 289 isolates from cases of human campylobacteriosis whose onset was within approximately 2 weeks from the date of processing. When there was genetic identity between broiler isolates and human isolates within the appropriate time frame, a retail product lot was classified as implicated in human disease. According to the results of this analysis, there were multiple clusters of human disease linked to the same process lot or lots. Implicated and nonimplicated retail product lots were compared for four lot descriptors: lot size, prevalence, mean contamination, and maximum contamination (as characterized by direct rinse plating). For retail product distributed fresh, Mann-Whitney U tests showed that implicated product lots had significantly (P = 0.0055) higher mean contamination than nonimplicated lots. The corresponding median values were 3.56 log CFU/carcass for implicated lots and 2.72 log CFU/carcass for nonimplicated lots. For frozen retail product, implicated lots were significantly (P = 0.0281) larger than nonimplicated lots. When the time frame was removed, retail product lots containing Campylobacter flaA SVR genotypes also seen in human disease had significantly higher mean and maximum contamination numbers than lots containing no genotypes seen in human disease for both fresh and frozen product. Our results suggest that cases of broiler-borne campylobacteriosis may occur in clusters and that the differences in mean contamination levels may provide a basis for regulatory action that is more specific than a presence-absence standard.

Prevalence of Encephalitozoon cuniculi Antibodies in Terrestrial Mammals in Iceland, 1986 to 1989

Antibodies to Encephalitozoon cuniculi were found in wild arctic foxes (Alopex lagopus), feral mink (Mustela vison), wood mice (Apodemus sylvaticus) and house mice (Mus musculus) in Iceland. Animals with antibodies were found throughout the country. No lesions attributable to encephalitozoonosis were found in adult animals necropsied. However, one arctic fox cub with a neurological disorder had pathological and serological evidence of encephalitozoonosis.

PARASITES OF THE ARCTIC FOX (ALOPEX LAGOPUS) IN ICELAND

Forty-four of 50 arctic foxes (Alopex lagopus) in Iceland harbored 15 species of intestinal parasites, including Protozoa: Eimeria sp. or Isospora sp. (in 4%); Trematoda: Cryptocotyle lingua (24%), Plagiorchis elegans (4%), Brachylaemus sp. (12%), Tristriata sp. (10%), and Spelotrema sp. (8%); Cestoda: Mesocestoides canislagopodis (72%), Schistocephalus solidus (2%), and Diphyllobothrium dendriticum (4%); Nematoda: Toxascaris leonina (50%), Toxocara canis (2%), Uncinaria stenocephala (4%), and eggs of the lung worm Capillaria aerophila (6%); and Acanthocephala: Polymorphus meyeri (8%) and Corynosoma hadweni (2%). Only four of the species previously had been recorded in Iceland. Eleven species are new records in Iceland and six appear to be new host records. Two additional nematodes, Stegophorus stercorarii and Syphacia sp., probably were ingested accidentally with the prey. Foxes from coastal habitats harbored 14 parasitic species while only five species were found in foxes from inland habitats. Arctic foxes from coastal habitats generally had higher helminth burdens and harbored more parasitic species per fox than foxes from inland habitats.

Combining two serological assays optimises sensitivity and specificity for the identification of Streptococcus equi subsp equi exposure

The detection of anti-Streptococcus equi antibodies in the blood serum of horses can assist with the identification of apparently healthy persistently infected carriers and the prevention of strangles outbreaks. The aim of the current study was to use genome sequencing data to develop an indirect enzyme linked immunosorbent assay (iELISA) that targets two S. equi-specific protein fragments. The sensitivity and specificity of the antigen A and antigen C iELISAs were compared to an SeM-based iELISA marketed by IDvet - diagnostic Vétérinaire (IDvet). Individually, each assay compromised specificity in order to achieve sufficient sensitivity (SeM iELISA had a sensitivity of 89.9%, but a specificity of only 77.0%) or sensitivity to achieve high specificity. However, combining the results of the antigen A and antigen C iELISAs permitted optimisation of both sensitivity (93.3%) and specificity (99.3%), providing a robust assay for the identification of horses exposed to S. equi.

Vaccinia virus complement control protein diminishes formation of atherosclerotic lesions: complement is centrally involved in atherosclerotic disease.

Complement is known to be activated in atherosclerotic lesions, but the importance of this event in disease pathology is a matter of debate. Studies of rabbits fed a high‐fat diet have indicated complement activation as a rate‐limiting step, whereas results from genetically modified mouse strains (ApoE−/− or LDLR−/−) have failed to support this finding. To resolve whether this reflects differences between species or between genetically driven and diet‐induced disease, we studied the effect of a complement inhibitor, vaccinia virus complement control protein (VCP), on C57BL/6 mice, the background strain of ApoE−/− and LDLR−/− mice. Atherosclerosis was induced by a high‐fat diet, and VCP (20 mg/kg) was injected once per week after the eighth week. Fatty streak development was monitored at 15 weeks by microscopic examination of oil red‐O‐stained sections from the root of the aorta. VCP injections led to significant (50%) reduction of lesion size (P= 0.004). Lesions were marked by gradual accumulation of lipids and macrophages but did not develop beyond the fatty streak stage. VCP activity disappeared from serum in 4 days, and the possibility therefore exists that a higher level of protection may be achieved by more frequent injections. We conclude that the development of fatty streaks in diet‐induced atherosclerotic disease can be significantly retarded by prophylactic treatment with a complement inhibitor. These results support previous findings from complement‐deficient rabbits and suggest that the pathogenesis of atherosclerosis in diet‐induced disease differs from that induced by major defects in lipid metabolism.

Prevalence and Genetic Relatedness of Antimicrobial-Resistant Escherichia coli Isolated From Animals, Foods and Humans in Iceland

The prevalence of resistant bacteria in food products in Iceland is unknown, and little is known of the prevalence in production animals. The aim of this study was to investigate the prevalence and genetic relatedness of antimicrobial‐resistant Escherichia coli from healthy pigs and broiler chicken, pork, broiler meat, slaughterhouse personnel and outpatients in Iceland. A total of 419 E. coli isolates were tested for antimicrobial susceptibility using a microbroth dilution method (VetMIC), and resistant strains were compared using pulsed‐field gel electrophoresis (PFGE). All samples were screened for enrofloxacin‐resistant strains with selective agar plates. The resistance rates among E. coli isolates were moderate to high from caecal and meat samples of pigs (54.1% and 28%), broilers (33.6% and 52%) and slaughterhouse personnel (39.1%), whereas isolates from outpatients showed moderate resistance rates (23.1%). Of notice was resistance to quinolones (minimum inhibitory concentrations: nalidixic acid ≥ 32, ciprofloxacin ≥ 0.12 and enrofloxacin ≥ 0.5), particularly among broiler and broiler meat isolates (18.2% and 36%), as there is no known antimicrobial selection pressure in the broiler production in Iceland. The majority (78.6%) of the resistant E. coli isolates was genotypically different, based on PFGE fingerprint analyses and clustering was limited. However, the same resistance pattern and pulsotype were found among isolates from broiler meat and a slaughterhouse worker, indicating spread of antimicrobial‐resistant E. coli from animals to humans. Diverse resistance patterns and pulsotypes suggest the presence of a large population of resistant E. coli in production animals in Iceland. This study gives baseline information on the prevalence of antimicrobial‐resistant E. coli from production animals, and their food products in Iceland and the moderate to high resistance rates emphasize the need for continuing surveillance. Further studies on the origin of the resistant strains and the genetic relatedness of strains of different origin are needed.

Olfactory Absorption of Insulin to the Brain

AbstractA vast number of potent neuropharmaceuticals, many of which are peptides, are excluded from entry into the brain because of the highly selective blood-brain barrier. The fact that a number of drugs have been shown to be transported directly to the central nervous system following application to the olfactory region of the nose is therefore of major interest. In the present study, the feasibility of delivering peptides to the brain via the olfactory route was assessed using insulin as a model peptide. Systemic hyperinsulinemia induced by subcutaneous injection did not significantly reduce the amount of 125I-insulin transported from the nose to the brain in vivo, which suggests that the impact of systemic absorption on drug transport is minimal. A linear relationship was seen between insulin accumulation in the brain and the dose applied, without any relevant saturation. Contrary to what was expected, both systemic and olfactory absorption of insulin was enhanced when the pH of the medium was near t...

Selective delivery of insulin into the brain: Intraolfactory absorption

Abstract The distribution of insulin between blood and brain was investigated in mice. The drug was administered subcutaneously and by instilling the drug to the olfactory region of the nasal cavity. The concentrations of insulin in the biological samples were measured by a γ-counter. A significantly higher concentration of insulin was measured in the brain, following intraolfactory administration compared to subcutaneous injection. The absorption was also found to be very rapid. Ten minutes after the administration the concentration in the brain had reached 193 counts/min per g (equivalent to 37 μU insulin/g tissue), which was 487%, higher than achieved after subcutaneous injection. The majority ( > 99%) of the CNS drug development programmes are devoted solely to CNS drug discovery and less than 1% are devoted to drug delivery. The results suggest that it may be possible to achieve absorption directly into the brain, by-passing the blood-brain barrier. The olfactory region may be the key for this absorption.