The phenology and population dynamics of Culicoides spp. in different ecosystems in The Netherlands

The Netherlands has enjoyed a relatively free state of vector-borne diseases of economic importance for more than one century. Emerging infectious diseases may change this situation, threatening the health of humans, domestic livestock and wildlife. In order to be prepared for the potential outbreak of vector-borne diseases, a study was undertaken to investigate the distribution and seasonal dynamics of candidate vectors of infectious diseases with emphasis on bluetongue vectors (Culicoides spp.). The study focused primarily on the relationship between characteristic ecosystems suitable for bluetongue vectors and climate, as well as on the phenology and population dynamics of these vectors.Twelve locations were selected, distributed over four distinct habitats: a wetland area, three riverine systems, four peat land areas and four livestock farms. Culicoides populations were sampled continuously using CO₂-baited counterflow traps from July 2005 until August 2006, with an interruption from November 2005 to March 2006. All vectors were identified to species level. Meteorological and environmental data were collected at each location.Culicoides species were found in all four different habitat types studied. Wetland areas and peat bogs were rich in Culicoides spp. The taxonomic groups Culicoides obsoletus (Meigen) and Culicoides pulicaris (Linnaeus) were strongly associated with farms. Eighty-eight percent of all Culicoides consisted of the taxon C. obsoletus/Culicoides scoticus. On the livestock farms, 3% of Culicoides existed of the alleged bluetongue vector Culicoides dewulfi Goetghebuer. Culicoides impunctatus Goetghebuer was strongly associated with wetland and peat bog. Many Culicoides species were found until late in the phenological season and their activity was strongly associated with climate throughout the year. High annual variations in population dynamics were observed within the same study areas, which were probably caused by annual variations in environmental conditions.The study demonstrates that candidate vectors of bluetongue virus are present in natural and livestock-farm habitats in the Netherlands, distributed widely across the country. Under favourable climatic conditions, following virus introduction, bluetongue can spread among livestock (cattle, sheep and goats), depending on the nature of the viral serotype. The question now arises whether the virus can survive the winter conditions in north-western Europe and whether measures can be taken that effectively halt further spread of the disease.     

Risk of infection with Leishmania spp. in the canine population in the Netherlands

The dog is the main reservoir of Leishmania infantum, the causative agent of visceral leishmaniasis (VL) in humans in Southern Europe. In order to identify the risk of dogs from a Leishmania non-endemic area traveling to a Leishmania-endemic area becoming infected and the risk of transmitting infection to humans in non-endemic areas an investigation was performed, in which the results of a questionnaire were combined with the results of a serologic survey. The questionnaire was sent to 1478 at random chosen families in the Netherlands. Of the 59.0% responders 28.0% had one or more dogs and 4.8% of these dogs had visited Southern Europe during the summer period of that year. On a total population of 1,200,000 dogs in the Netherlands, this means that each year some 58,000 dogs are at risk of being exposed to a Leishmania infection in Southern Europe. During the period 1990-1992 blood was collected for serology in 1911 dogs presented to the Utrecht University Clinic because of clinical problems not related to leishmaniasis, of which 434 had been in Southern Europe in the foregoing years. None was serologically positive. From these data it can be deduced that the highest chance to obtain leishmaniasis during a vacation in Southern Europe is mathematically less than 1/434 or less than 0.23%. Serology was also performed during the period 1989-1993 in 597 dogs that had been in Southern Europe and were suspected of leishmaniasis. Titers were positive in 145 of these samples. Sixty-four of these dogs were born in the Mediterranean and had been imported into the Netherlands. Excluding these imported dogs, it was calculated that at least 0.027% of the 58,000 dogs yearly taken to Southern Europe during holidays become infected with Leishmania. In order to establish the risk of disease transmission for people in close contact with an infected dog, serum samples of owners and house mates of 37 dogs with leishmaniasis were tested. All 112 sera tested negative. It was concluded that the risk to get leishmaniasis was between 0.027% and 0.23% for the dog when taken to Southern Europe during vacation, and that the risk for owners in non-endemic areas to get leishmaniasis from an infected dog is minimal.     

Culicoides species associated with sheep in the Netherlands and the effect of a permethrin insecticide

Culicoides spp. act as vectors for a number of viral diseases of animals including bluetongue in sheep. The aims of this study were to determine: (1) which Culicoides spp. are associated with sheep in The Netherlands; (2) the time of the day when they are most active; and (3) the effect of treatment of animals with a permethrin insecticide. Two pairs of sheep were each housed within mosquito tents of either one or two layers of netting and all trapped Culicoides spp. were identified microscopically. For the permethrin insecticide study, one of two pairs of sheep was treated with 3.6% permethrin and all animals were housed in tents of similar design. Of the 6210 midges captured, 54.1% were identified as C. chiopterus and 42.7% as C. obsoletus. C. imicola was not identified. The average insect feeding rate was 35-40% and midge activity was greatest around sunset. Permethrin treatment reduced the number of midges captured by 50% and also resulted in a decrease in the percentage of midges that had fed. The findings provide useful information on the behaviour and distribution of Culicoides spp. that will facilitate the development of appropriate control strategies to minimise the risk of insect-vector borne virus diseases such as bluetongue.     

Molecular detection of Culicoides spp. and Culicoides imicola, the principal vector of bluetongue (BT) and African horse sickness (AHS) in Africa and Europe

Bluetongue (BT) and African Horse Sickness (AHS) are infectious arthropod-borne viral diseases affecting ruminants and horses, respectively. Culicoides imicola Kieffer, 1913, a biting midge, is the principal vector of these livestock diseases in Africa and Europe. Recently bluetongue disease has re-emerged in the Mediterranean Basin and has had a devastating effect on the sheep industry in Italy and on the islands of Sicily, Sardinia, Corsica and the Balearics, but fortunately, has not penetrated onto mainland France and Spain. To survey for the presence of C. imicola, an extensive light-trap network for the collection of Culicoides, was implemented in 2002 in southern mainland France. The morphological identification of Culicoides can be both tedious and time-consuming because its size ranges from 1.5 to 3 mm. Therefore, an ITS1 rDNA polymerase chain reaction (PCR)-based diagnostic assay was developed to rapidly and reliably identify Culicoides spp. and C. imicola. The aim of this work was to set up a rapid test for the detection of C. imicola amongst a pool of insects collected in areas at risk for BT. The sequence similarity of the rDNA (nuclear ribosomal DNA), which is greater within species than between species, is the foundation of its utilisation in species-diagnostic assays. The alignment of the 11 ITS1 sequences of Culicoides obtained from Genbank and EMBL databases helped us to identify one region in the 5' end and one in the 3' end that appear highly conserved. PCR primers were designed within these regions to amplify genus-specific fragments. In order to set up a C. imicola-specific PCR, another forward primer was designed and used in combination with the previously designed reverse primer. These primers proved to be highly specific and sensitive and permitted a rapid diagnostic separation of C. imicola from Culicoides spp.     

An unrecognized species of the Culicoides obsoletus complex feeding on livestock in the Netherlands

In studies on Culicoides attacking livestock in the Netherlands, we chanced upon a species of the Obsoletus complex that we do not recognize, but whose dark wing pattern is distinctive. Nine cytochrome c oxidase (CO1) sequences of our so-called ‘dark obsoletus’ support its status as a separate species, the sequences differing significantly from those representing Culicoides obsoletus (Meigen) (90–91% homology) and Culicoides scoticus Downes & Kettle (87–88% homology). In the last decade, several research groups in Europe have encountered ‘mystery species’ related to C. obsoletus and in some instances have made their sequences for various genetic loci available in GenBank. These include a CO1 series submitted from Sweden in 2012 (annotated as ‘obsoletus 01, 02, or 03 MA-2012′) and of which some share a 99% identity with our sequences for ‘dark obsoletus’. Without doubt, the series from the Netherlands, along with a portion of the Swedish submissions, together represent a single species (‘dark obsoletus’). Whether this species is referable to the Russian Culicoides gornostaevae Mirzaeva recorded recently from Norway, Sweden and Poland, and based solely upon the external morphology of the male, is not clear. The presence in Western Europe of multiple undescribed species related to C. obsoletus means that the taxonomy of this important vector complex is not fully resolved; consequently, we know little about these cryptic species with regard to seasonality, geographic range and host preference. This is undesirable given that Culicoides-borne arboviruses causing disease in livestock are moving more regularly out of the tropics and spreading north into temperate latitudes.     

The importance of different pig diseases in the Netherlands

As part of the project 'Clean pigs', IPG Institute for Pig Genetics BV made an inventory of the impact of different diseases on the Dutch pig industry. An expert panel assessed the importance of the different diseases with regard to public health, farm economy, the pig sector and export sales. The possibilities for the diagnosis of the different pathogens were listed and the goal for the next years, was set. Diseases were classified into three categories, according to their importance for the Dutch pig industry (Table 2). The diseases that can be eradicated are Salmonella (only specific strains), Pasteurella multocida DNT + PAR), Actinobacillus pleuropneumoniae, Haematopinus suis (lice) and Sarcoptes scabei (var. suis) (mange). National introduction of list A en most list B diseases of the OIE classification must be prevented.     

The population dynamics of nematode parasites of sheep in northern England.

An investigation into the population dynamics of sheep nematodes was carried out between April 1970 and September 1971. Faecal egg counts were made from both ewes and lambs at weeks intervals while grass samples were taken fortnightly throughout the summer and monthly during the winter. From these results absolute numbers of nematode eggs and infective larvae were estimated and these figures used as a basis for determining the level of mortality occurring during development from egg to infective larva. The figures were also used to assess the relative importance of the ewe 'spring rise' and the egg output of the lambs in June while grazing contaminated pasture. These calculations suggest that larval mortality was very high and that it may vary considerably from year to year depending upon the climatic conditions prevailing. There was also an indication that the ewe 'spring rise' was the major source of the wave of infection which occurs in the lambs in late August and September.     

Culicoides spp. (Diptera: Ceratopogonidae) Associated with Farmed Ostriches (Struthio camelus) in Botswana

Veterinary Research Communications -     

Endophily in Culicoides associated with BTV-infected cattle in the province of Limburg, south-eastern Netherlands, 2006

Culicoides were captured at a BTV-infected dairy near Gulpen in the province of Limburg (south-east Netherlands) between 14 September and 4 October 2006. Onderstepoort-type blacklight traps were used to sample Culicoides both inside and outside a partially open shed housing 11 cattle. A total of 28 light trap collections were made at the shed and yielded: In Europe, the blacklight trap is used widely for the nocturnal monitoring of Culicoides; a drawback to this approach is that this trap cannot be used to sample midges that are active during the day. Because diurnal biting in vector Culicoides may constitute a significant and underestimated component of BTV transmission a novel capture methodology will be required in future and is discussed briefly.     

The occurrence of Campylobacter spp. and Salmonella spp. in gulls in northern Germany

Out of a total of 207 gulls--20 Black headed gulls, 185 Herring gulls and 2 Common gulls--128 (62%) and 23 (11%) birds, respectively, were infected with Campylobacter spp. (C.) and Salmonella spp. C. jejuni was predominant in gulls less than two years old (89%) and C. coli in older birds (75%). Furthermore, the infection rate with Campylobacters was depending on the habitats of the birds. The rate for C. jejuni and C. coli, respectively, was in gulls from regional garbage dumps 78% and 4%, from the coast 58% and 21%, and from islands 47% and 47% of the isolations in the corresponding area. Salmonellae were mainly isolated in the period from September to February from gulls less than one year old and from birds from the coast.     

The prevalences of Brachyspira spp. and Lawsonia intracellularis in Swedish piglet producing herds and wild boar population

The aim of the present study was to survey the prevalences of the enteric pathogens Brachyspira hyodysenteriae, Brachyspira pilosicoli and Lawsonia intracellularis in Swedish growing pigs and in the Swedish wild boar population and to relate these findings to clinical signs. The study included 105 randomly selected herds, constituting approximately one third of Swedish herds with a herd size of >100 sows. The herds were located all over the country. In these herds, growth promoters were not used and pigs sampled were not subjected to any medication. From each herd, samples were taken from 10 growing pigs aged 8-12 weeks, corresponding to approximately 2.5% of all growing pigs present in the herd at the sampling occasion. If possible, the samples were taken from pigs with diarrhoea. Forty-eight faecal samples and 71 rectal swabs were also taken from free-living wild boars (31 piglets, 19 growers and 21 adult animals) at shooting. The samples were analysed by culture and biochemical tests for the presence of Brachyspira spp. and by nested PCR for the presence of L. intracellularis. Brachyspira hyodysenteriae was not demonstrated in any sample. Brachyspira intermedia was detected in 22 samples originating from 15 herds, Brachyspira innocens/Brachyspira murdochii was detected in 370 samples from 82 herds and B. pilosicoli was detected in 134 samples originating from 34 herds. In 21 herds and in 534 samples, no Brachyspira spp. were detected. Lawsonia intracellularis was demonstrated in 285 samples from 50 herds. Further, 418 samples from conventional herds were negative with respect to L. intracellularis and in 345 samples the PCR had been inhibited. All samples from the wild boars were negative for Brachyspira spp., 12 of 48 samples were negative for L. intracellularis, and in 36 wild boar samples, the PCR was inhibited.     

Abundance of biting midge species (Diptera: Ceratopogonidae,Culicoides spp.) on cattle farms in Korea

Culicoides biting midges were collected on three cattle farms weekly using light traps overnight from May to October between 2010 and 2011 in the southern part of Korea. The seasonal and geographical abundance of Culicodes spp. were measured. A total of 16,538 biting midges were collected from 2010 to 2011, including seven species of Culicoides, four of which represented 98.42% of the collected specimens. These four species were Culicodes (C.) punctatus (n = 14,413), C. arakawae (n = 1,120), C. oxystoma (n = 427), and C. maculatus (n = 318). C. punctatus was the predominant species (87.15%).     

Seasonal abundance of four Culicoides spp. (Diptera: Ceratopogonidae) at Al-Ahsa oasis,Eastern Province,Saudi Arabia

This report constitutes the first study of Culicoides spp. and their seasonal abundance at Al-Ahsa, the largest oasis in the Eastern Province of Saudi Arabia. New Jersey light traps were used to collect the midges at Mastock farm and Al-Mansura village. The mean monthly abundance was determined from October 1993 to October 1994. The mean monthly number per trap reached its minimum value during January 1994, increasing gradually from February to reach its maximum value during September 1994. During the study period, the following species were collected: Culicoides schultzei group (September), non-spotted group of Culicoides (September), Culicoides imicola (May) and Culicoides newstaedi (March). The potential importance of the Culicoides spp. in relation to arboviral activity in Saudi Arabia is discussed.     

白喉乌头种群生长动态初步研究

白喉乌头(Aconitum leucostomum)是新疆伊犁河谷山地草甸草地广泛分布的多年生有毒植物,严重危害着当地草原畜牧业的发展。为掌握其生长发育状态,为合理防治提供科学依据,对白喉乌头从返青到开花结实期(5月3日-7月17日)的高度(H)、密度(D)、株丛径(Z)和地上生物量(W)等每15d测定一次,监测其生长动态,并采用一元非线性拟合的方法,对白喉乌头的生长动态进行拟合,并对高度、密度、株丛径与地上生物量的关系进行了逐步回归分析。研究结果表明:白喉乌头的高度、株丛径和地上生物量随时间的增加而增大,其增长模型分别是:幂函数模型、Long-modified模型和幂函数模型,其拟合方程分别是H=4.6978T0.513551、Z=(494.1776+231.5106T)0.339913和W=2.6393T0.781408;白喉乌头的密度先增大后减小,符合房室模型,其方程式为D=114.7676e(-0.005049T)-100.8592e(-0.095765T);在同一时间高度、株丛径与地上生物量之间呈显著相关的回归关系:W=39.7814364+5.429432775H-7.108936940Z;高度是影响白喉乌头地上生物量最重要的因素。因此,可以通过控制高度来控制白喉乌头的繁殖。     

青藏高原祁连山东段高寒小嵩草草甸植物种群物候学研究(简报)

通过对青藏高原祁连山东段高寒小嵩草草甸主要植物种群物候特性的观测研究,采用聚类分析和主分量排序法将小嵩草草甸26种植物种群划分为不同的物候类群。利用相关系数排序法指出了影响不同物候期的主要生态因子依次是温度、降水量和日照时数。研究结果对物候资料积累、分析以及放牧管理具有指导意义。     

Efficacy of different dosages of ivermectin injectable against the Hypoderma spp. in yaks

A chemotherapy trial was conducted to determine the lowest dosage of injectable preparation of ivermectin against Hypoderma spp. infestation in yaks in Tibetan areas in Tianzhu county, Gansu province, in northwest of China. One hundred and sixty yaks were randomly divided into four groups of 40 yaks for the trial. The first three groups were treated by subcutaneous injection in the neck with 0.1% ivermectin (respectively, 1, 5, 10 microg/kg body weight). The fourth group was not treated and considered as control group. All the experiments were performed in November 2000 and the animals were examined for the presence of warbles in the next March and May. The results indicated that there was no warbles found on the back of treated animal while third stage larvae were palpated on back of some of the yaks in control group. It is concluded that dosage of 1 microg/kg ivermectin injectable was sufficient to kill or stop development of larvae of Hypoderma spp. in naturally infected yaks if administrated in November.     

Comparison of black and white light for collecting Culicoides imicola and other livestock-associated Culicoides species in South Africa

Comparison of the effectiveness of 8W fluorescent black and white light sources, in two 4x4 Latin squares (16 replicates) designs under South African conditions, showed black light to be up to three time more effective in collecting Culicoides imicola Kieffer (Diptera, Ceratopogonidae) and other South African Culicoides species. Four Culicoides species, which were collected in low numbers with black light, were not collected in traps equipped with the white light source. No significant difference was found in the parous rate of the C. imicola populations as determined by the two light sources. The study highlighted the superiority of black light as a preferred collection method for C. imicola, considered to be the most widespread and abundant vector of livestock orbiviruses. The results underline the need to develop and adopt standard techniques for measuring the variables of vectorial capacity.     

The effect of a probiotic Enterococcus faecium product in diets of healthy dogs on bacteriological counts of Salmonella spp., Campylobacter spp. and Clostridium spp. in faeces.

An experiment was conducted to determine the effects of a probiotic on selected faecal bacteria of healthy dogs under different feed and environmental conditions. For the study 12 dogs kept in households were used for an 18-day supplementation with a patented commercially available strain of E. faecium NCIB 10415 (Enteroferm). In order to minimize losses the probiotic product was orally applicated once a day before meals at a dose of 2 g per dog (9.2 x 10(9) CFU). The faeces were collected before the beginning of the supplementation and at the end of the 18-day application period. In order to exclude contamination, all faeces were taken rectally. Before and at the end of the experimental period total Salmonella spp., Campylobacter spp. and Clostridium spp. counts were determined in fresh faeces using selective media. It was demonstrated that the 18-day application of the probiotic E. faecium product induced modifications on the gastrointestinal microflora in all dogs. While Salmonella spp. and Campylobacter spp. counts were in majority of the dogs higher than before the application. Clostridium spp. counts were significantly reduced in 10 of 12 dogs. According to the guidelines for the evaluation of the efficiency of microorganisms in dogs a relevant efficacy effect was supported by this data. However, a beneficial effect of the probiotic product on healthy dogs remains questionable.