Henrik Skovgård

Flies and Campylobacter Infection of Broiler Flocks

A total of 8.2% of flies caught outside a broiler house in Denmark had the potential to transmit Campylobacter jejuni to chickens, and hundreds of flies per day passed through the ventilation system into the broiler house. Our study suggests that flies may be an important source of Campylobacter infection of broiler flocks in summer.

Use of Fly Screens to Reduce Campylobacter spp. Introduction in Broiler Houses

Fly screens that prevented influx of flies in 20 broiler houses during the summer of 2006 in Denmark caused a decrease in Campylobacter spp.-positive flocks from 51.4% in control houses to 15.4% in case houses. A proportional reduction in the incidence of chicken-borne campylobacteriosis can be expected by comprehensive intervention against flies in broiler production houses.

Influxed Insects as Vectors for Campylobacter jejuni and Campylobacter coli in Danish Broiler Houses

The vector potential of flies (Diptera: Brachycera) for spread of Campylobacter jejuni and Campylobacter coli on 5 Danish broiler farms was evaluated in a longitudinal field study from April to November 2004. First, the prevalence of C. jejuni- and C. coli-positive flies was determined in 2,816 flies captured from farm surroundings. Each individual fly was macerated, preenriched in Bolton broth for 24 h at 42 degrees C, streaked onto modified Campylobater blood-free selective agar and incubated under microaerobic conditions for 48 h at 42 degrees C. Second, the influx of insects to broiler houses was estimated by trapping of insects (n = 5,936) in ventilation vents. In total, 31 flies (28 of which were of the Muscidae family) caught in farm surroundings were Campylobacter spp.-positive (C. jejuni, n = 7; C. coli, n = 23; other Campylobacter spp., n = 1). Musca domestica (L) (house fly) was more frequently (P < 0.0376) positive than other fly species collected. Other positive fly species were Stomoxys calcitrans (L) (stable fly; n = 4), Muscina stabulans (Fallén) (false stable fly; n = 1), Mesembrina meridiana (L) (noon fly; n = 1) and Hydrotaea sp. (black dump fly; n = 1). One Lucilia caesar (L) (green bottle fly) of the Calliphoridae family and 2 flies of unidentified species were also positive. The prevalence of Campylobacter spp.-positive flies varied from 0.0 in April to a peak of 16.3% in July and decreasing to 2.0% in October on a farm with pig production. On 4 broiler farms without other livestock, the prevalence was constantly below 1.0%. The average influx of insects per broiler rotation was estimated to be 30,728 +/- 2,443 SE (range 2,233 to 180,300), of which 21.4% were flies. The influx of insects correlated with the flow (m(3)/h) of ventilation air (P < 0.0078) and with the outdoor temperature (P < 0.0032). We conclude that the influx of large numbers of flies to broiler houses constitutes a considerable risk for infection of broilers with C. jejuni and C. coli.

Molecular identification of bloodmeals from biting midges (Diptera: Ceratopogonidae: Culicoides Latreille) in Denmark

Culicoides vectors are critical to the survival and transmission of bluetongue virus as infection only occurs in areas or regions where competent vectors are present. The success of Culicoides biting midges as vectors is mainly related to their vast population sizes and to their means of dispersal. Their choice of host for blood feeding is sparsely described. The aim of the present study was to establish methods for the identification of bloodmeal hosts and determine the identity and diversity of bloodmeals of vertebrate hosts from wild-caught biting midges near livestock farms. The study includes some of the most common and abundant species of biting midges in Denmark: Culicoides obsoletus, Culicoides scoticus, Culicoides pulicaris and Culicoides punctatus. We collected 8,378 biting midges including nine species of Culicoides of which blood-fed specimens were found from six species. We identified 251 blood engorged biting midges, and hosts were identified in 115 of 125 analysed specimens (90%). Cow, roe deer, horse, mallard and wood pigeon were identified as hosts. The most abundant host species was cow, which constituted 73.9% of the total identified bloodmeals, but the common wood pigeon was found with a frequency as high as 18.3%. In conclusion, the molecular methods applied were proven useful in identifying bloodmeal hosts from different Culicoides species. The results indicate that Culicoides species are opportunistic in their choice of bloodmeal host with a preference for cattle when present, which is important to have in mind for epidemiologist when making predictive models. Accordingly, the results of this study will add useful parameters for modelling bluetongue virus transmission and in the development of veterinary contingency plans.

Life Table Study of Sitotroga cerealella (Lepidoptera: Gelichiidae), a Strain from West Africa

Life table studies for the Angoumois grain moth, Sitotroga cerealella (Olivier), a pest on stored maize, Zea mays L., in West Africa, were conducted as part of the expansion of a mathematical simulation model that has been developed for two pests of stored maize. The effects of four temperatures (20, 25, 30, and 35 degrees C) and two relative humidity levels (44 and 80%) on developmental time, age-specific survivorship and fecundity, sex ratio, and intrinsic rate of natural increase (r(m)) of S. cerealella were investigated. Sex ratio was close to 1:1 at all temperatures and humidity. Minimum development time occurred close to 32 degrees C and 80% RH for both males and females, and developmental time of females was significantly shorter than that of males. Immature survivorship was highest between 25 and 30 degrees C and 80% RH and lowest at 35 degrees C under both humidity conditions. A similar low level was found at 20 degrees C and 44% RH. The greatest fecundity (124 eggs per female) occurred at 20 degrees C, 80% RH. The maximum r(m) value was 0.086 d(-1) at 30 degrees C and 80% RH, but the growth rate declined dramatically at 35 degrees C. If compared with the few other life table studies conducted on this species on maize in India and North America, some variation among the strains becomes evident. A common conclusion for the current study and previous ones is that optimal population development for S. cerealella occurs at approximately 30 degrees C and at high humidity.

Molecular differentiation of Culicoides biting midges (Diptera: Ceratopogonidae) from the subgenus Culicoides Latreille in Denmark

Identification of Culicoides biting midges to species has attracted attention due to the recent outbreak of bluetongue disease in Northern Europe. Identification of Culicoides to species level has been based on morphological characters and is difficult as several species belonging to species complexes are hard to distinguish. We evaluated the use of the mitochondrial DNA cytochrome oxidase I gene (COI) barcode region in the identification of species within the subgenus Culicoides. COI barcode sequence divergence within species was <1%, whereas it ranged from 12.5% to 19.8% between subgenus Culicoides species. The divergence of subgenus Culicoides species to C. nubeculosus from the subgenus Monoculicoides ranged from 24.4% to 26.1%. Specimens were differentiated into eight unique clusters, including the four common Palaearctic species Culicoides punctatus, Culicoides pulicaris, Culicoides impunctatus, and Culicoides grisescens. Additionally, this study confirms the existence of Culicoides halophilus as a valid taxon and presents the first Culicoides deltus barcode sequences. Three additional groups of specimens were identified: Culicoides dk1 with a COI barcode diverging by 14.3% to 17.2% from other subgenus Culicoides species and Culicoides Kalix and Culicoides dk3, which diverged by 5.9% from each other and showed 12.5% to 17.6% divergence in COI barcode to subgenus Culicoides specimens.

Persistence of Low-Pathogenic Avian Influenza H5N7 and H7N1 Subtypes in House Flies (Diptera: Muscidae)

ABSTRACT Avian influenza caused by avian influenza virus (AIV) has a negative impact on poultry production. Low-pathogenic AIV (LPAIV) is naturally present in wild birds, and the introduction of the virus into domestic poultry is assumed to occur through contact with wild birds and by human activity, including the movement of live and dead poultry, and fomites such as clothing and vehicles. At present, the possible role of insects in the spread of AIV is dubious. The objective of the present work was to investigate the potential transmission of LPAIV by persistence of the virus in the alimentary tract of house flies, Musca domestica L. (Diptera: Muscidae). Flies were fed three virus concentrations of two AIV strains and then incubated at different temperatures for up to 24 h. The persistence of the two virus strains in the flies declined with increasing incubation temperatures and incubation periods. Similarly, increased virus uptake by the flies increased the persistence of virus. Persistence of infective AIV in flies differed significantly between the two virus strains. The laboratory experiments of the present study indicate that the house fly can be a potential carrier of AIV.

The Effects of Sex-Ratio and Density on Locomotor Activity in the House Fly, Musca domestica

Abstract Although locomotor activity is involved in almost all behavioral traits, there is a lack of knowledge on what factors affect it. This study examined the effects of sex—ratio and density on the circadian rhythm of locomotor activity of adult Musca domestica L. (Diptera: Muscidae) using an infra—red light system. Sex—ratio significantly affected locomotor activity, increasing with the percentage of males in the vials. In accordance with other studies, males were more active than females, but the circadian rhythm of the two sexes was not constant over time and changed during the light period. There was also an effect of density on locomotor activity, where males at intermediate densities showed higher activity. Further, the predictability of the locomotor activity, estimated as the degree of autocorrelation of the activity data, increased with the number of males present in the vials both with and without the presence of females. Overall, this study demonstrates that locomotor activity in M. domestica is affected by sex—ratio and density. Furthermore, the predictability of locomotor activity is affected by both sex—ratio, density, and circadian rhythm. These results add to our understanding of the behavioral interactions between houseflies and highlight the importance of these factors when designing behavioral experiments using M. domestica.

Retention of Campylobacter (Campylobacterales: Campylobacteraceae) in the house fly (Diptera: Muscidae).

ABSTRACT The house fly (Musca domestica L.) may transmit Campylobacter to broiler flocks. We assessed the retention time of house flies for Campylobacter jejuni at five temperatures and three doses. Flies were inoculated individually at their proboscis with 1.6 × 107 CFU (colony forming units) of C. jejuni and incubated at 15, 20, 25, 30, and 35°C. Furthermore, a dose experiment was conducted at 25°C where individual flies were inoculated in three series: 6.5 × 106, 6.0 × 104, and 8.2 × 102 C. jejuni CFU. Whole flies were tested for C. jejuni carriage at 0, 6, 12, 18, and 24 h by initial preenrichment in Bolton broth, which afterwards was streaked on modified mCCDA agar plates and incubated under micro-aerobic conditions. The results showed that the time C. jejuni remained in flies declined over time with ascending temperatures and when reducing the inoculation dose. All flies stayed Campylobacter positive 24 h postinoculation at 15°C whereas only one-third of the flies were positive at 20°C and few to none at 25, 30, and 35°C. When combinations of temperature and retention time were expressed as accumulated day-degrees, data could be adequately fitted using a generalized linear mixed model that included a linear effect of day-degrees and the difference between the lowest and the two highest doses. Based on model predictions of selected combinations of temperature and dose, the time for 50% and 1% of flies containing Campylobacter was calculated. It is suggested that house flies are mainly short distance carriers of C. jejuni.

The effects of temperature and innate immunity on transmission of Campylobacter jejuni (Campylobacterales: Campylobacteraceae) between life stages of musca domestica (Diptera: Muscidae)

ABSTRACT The house fly (Musca domestica L.) is a well-established vector of human pathogens, including Campylobacter spp., which can cause infection of broiler chicken flocks, and through contaminated broiler meat can cause outbreaks of campylobacteriosis in humans. We investigated whether Campylobacter jejuni (Jones) could be transferred between life stages of M. domestica (larvae-pupae-adults) and determined bacterial counts of C. jejuni at different time points after bacterial exposure. C. jejuni was transmitted from infected larvae to pupae, but not to the adult stage. Infected larvae maintained at 25°C had mean bacterial numbers of 6.5 ± 0.2 SE log10 (colony forming units [CFU]/g) that subsequently dropped to 3.6 ± 0.3 SE log10 (CFU/g) 8 h after infection. Pupae originating from infected larvae contained mean bacterial numbers of 5.3 ± 0.1 SE log10 (CFU/g), and these numbers dropped to 4.8 ± 0.1 SE log10 (CFU/g) 24 h after pupation. The decline in C. jejuni numbers during pupal development coincided with increased expression of antimicrobial peptides, including cecropin, diptericin, attacin, and defensin, in the larva-pupa transition stage and a later second peak in older pupae (4 or 48 h). Conversely, there was a reduced expression of the digestive enzyme, lysozyme, in pupae and adults compared with larvae.