Specie di Culicoides (Diptera: Ceratopogonidae) vettori del virus della Bluetongue in Sud Africa

The aim of this paper is to consolidate vector competence studies on Culicoides midges (Diptera: Ceratopogonidae) as vectors of Bluetongue virus (BTV) done over a period 25 years at the Agriculture Research Council ‑ Onderstepoort Veterinary Institute in South Africa. In 1944, it was demonstrated for the first time in South Africa that Culicoides midges transmit BTV. In 1991, field‑collected Culicoides imicola were fed on blood containing BTV‑3 or BTV‑6 and the infection rates were established as being 31% and 24%, respectively. In 1998, Culicoides bolitinos was shown to have a higher infection prevalence and virus titre/midge than C. imicola. This species was then shown to have a higher transmission potential for BTV‑1 over a range of incubation temperatures wider than the one showed by C. imicola. Attenuation of BTV also does not reduce its ability to infect competent Culicoides species. Oral susceptibility studies, involving 29 BTV isolates of various serotypes, indicated differences between various geographic virus isolates and Culicoides populations evaluated. While low recovery rates of European BTV strains from South African Culicoides species suggest co‑adaptation between orbiviruses and vectors in a given locality, co‑adaption was shown not to be essential for virus transmission. Cumulative results since 1991 provide evidence that at least 13 livestock‑associated Culicoides species are susceptible to BTV. Susceptibility results are supported by field isolations from 5 of these species. This implies that multi‑vector potential for the transmission of BTV will complicate the epidemiology of BT. It must be emphasised that neither oral susceptibility nor virus isolation/ detection from field‑collected specimens is proof that a species is a confirmed field vector.Il presente lavoro ha l’obiettivo di descrivere i risultati degli ultimi 25 anni di ricerca condotta all’Agriculture Research Council ‑ Onderstepoort Veterinary Institute in Sud Africa, sul ruolo dei Culicoides (Ditteri: Ceratopogonidae) come vettori del virus della Bluetongue (BTV). Nel 1944, in Sud Africa, è stato dimostrato per la prima volta il loro ruolo di vettori di BTV. Nel 1991, Culicoides imicola catturati in campo e nutriti con sangue contenente i sierotipi 3 o 6 del BTV hanno evidenziato percentuali di infezione, rispettivamente, del 31% e 24%. Nel 1998 è stato dimostrato che Culicoides bolitinos ha una prevalenza di infezione e un titolo virale per insetto superiori a quelli di C. imicola. È stato quindi dimostrato che C. bolitinos ha un maggior potenziale di trasmissione di BTV-1 in un intervallo di temperatura di incubazione più esteso di quello riportato per C. imicola. L’attenuazione del BTV non riduce la sua capacità di infettare specie competenti di Culicoides. Studi di competenza vettoriale attraverso infezione orale con 29 ceppi diversi appartenenti a vari sierotipi di BTV hanno evidenziato differenze a seconda dell'origine geografica dei ceppi e delle popolazioni di Culicoides esaminate. I bassi livelli di infezione dei ceppi europei di BTV in specie sudafricane di Culicoides suggeriscono l'esistenza di un possibile co-adattamento tra Orbivirus e vettori in una data zona, co-adattamento che tuttavia non è essenziale per la trasmissione del virus. In tutti questi anni è stato possibile dimostrare che almeno 13 specie di Culicoides associate al bestiame sono suscettibili al BTV. Tali risultati sono supportati da isolamenti di campo su 5 di queste specie. Ciò implica che il potenziale multi-vettoriale per la trasmissione di BTV complicherà l'epidemiologia di BT. Va sottolineato che né la suscettibilità orale né l’isolamento/rilevamento del virus da insetti catturati in campo provano che una data specie sia vettore certo del BTV.http://www.izs.it/vet_italiana/issues_vet_it.htmam2016Veterinary Tropical Disease

An investigation of culicoides (Diptera: Ceratopogonidae) as potential vectors of medically and veterinary important arboviruses in South Africa

Culicoides-borne viruses such as bluetongue, African horse sickness, and Schmallenberg virus cause major economic burdens due to animal outbreaks in Africa and their emergence in Europe and Asia. However, little is known about the role of Culicoides as vectors for zoonotic arboviruses. In this study, we identify both veterinary and zoonotic arboviruses in pools of Culicoides biting midges in South Africa, during 2012–2017. Midges were collected at six surveillance sites in three provinces and screened for Alphavirs, Flavivirus, Orthobunyavirus, and Phlebovirus genera; equine encephalosis virus (EEV); and Rhaboviridae, by reverse transcription polymerase chain reaction. In total, 66/331 (minimum infection rate (MIR) = 0.4) pools tested positive for one or more arbovirus. Orthobunyaviruses, including Shuni virus (MIR = 0.1) and EEV (MIR = 0.2) were more readily detected, while only 2/66 (MIR = 0.1) Middelburg virus and 4/66 unknown Rhabdoviridae viruses (MIR = 0.0) were detected. This study suggests Culicoides as potential vectors of both veterinary and zoonotic arboviruses detected in disease outbreaks in Africa, which may contribute to the emergence of these viruses to new regionsA US CDC Global Disease Detection grant for Zoonotic arboviruses, the NRF, the Meat Industry Trust and the Poliomyelitis Research foundation.https://medicalpapers.cn1699.com/journal/viruses-baselpm2022Medical Virolog

The prevalence of Culicoides spp.in 3 geographic areas of South Africa

The seasonal abundance of Culicoides midges, the vector of Bluetongue and African horse sickness viruses (BTV/AHSV) and the presence of viruses in midges were determined in 3 geographic areas in South Africa. In the Onderstepoort area, more than 500,000 Culicoides midges belonging to 27 species were collected. Eighteen midge species were collected throughout Winter and the presence of AHSV and BTV RNA in midges was detected using real time reverse transcription quantitative polymerase chain reaction. The nucleic acid of AHSV was found in 12 pools out of total pools of 35 Culicoides. Twenty‑five Culicoides species were detected in the Mnisi area. The RNA of BTV was detected in 75.9% of the midge pools collected during Winter and 51.2% of those collected during Autumn. Antibodies for BTV were detected in 95% of cattle sampled using a competitive enzyme‑linked immunosorbent assay (cELISA). The dominant species in these 2 areas was Culicoides imicola. Eight Culicoides species were collected in Namaqualand. Culicoides imicola represented the 0.9% and Culicoides bolitinos the 1.5% of total catches, respectively. Antibodies for AHSV were detected in 4.4% of 874 equines tested using an indirect ELISA. Results showed that transmission of AHSV and BTV can carry on throughout Winter and the outbreak may begin as soon as Culicoides populations reach a certain critical level. [Abstract] I Culicoides sono noti vettori del virus della Bluetongue (BTV) e del virus della Peste equina africana (AHSV). Il lavoro riporta i risultati sull'abbondanza stagionale di Culicoides e sulla presenza di BTV e AHSV in diversi vettori, in tre aree geografiche del Sudafrica. Nell'area di Onderstepoort sono stati individuati più di 500.000 esemplari di Culicoides appartenenti a 27 specie diverse. Durante la stagione invernale sono state individuate 18 specie. Frammenti di RNA di AHSV e BTV sono stati rilevati mediante specifiche RT-qPCR. L'RNA del virus AHSV è stato individuato in 12 pool di Culicoides su 35 esaminati. Nell'area di Mnisi sono state identificate 25 specie di Culicoides. L'RNA di BTV è stato rilevato nel 75,9% dei pool di Culicoides catturati durante la stagione invernale e nel 51,2% di quelli catturati durante la stagione autunnale. Anticorpi contro BTV sono stati osservati nel 95% dei sieri prelevati da bovini dell'area di Mnisi ed esaminati con il metodo c‑ELISA. La specie dominante in queste due aree è risultata Culicoides imicola. Nella regione di Namaqualand sono state individuate otto specie, C. imicola ha rappresentato lo 0,9% delle specie catturate e Culicoides bolitinos l'1,5%. Anticorpi contro il virus AHSV sono stati individuati mediante ELISA nel 4,4% degli 874 equini esaminati. I risultati ottenuti hanno dimostrato che in Sudafrica la trasmissione di BTV e AHSV può continuare durante la stagione invernale mentre, con ogni probabilità, si ha la comparsa dei primi focolai quando le diverse popolazioni di Culicoides raggiungono un livello riproduttivo critico

Culicoides species abundance and potential overwintering of African horse sickness virus in the Onderstepoort area, Gauteng, South Africa

In South Africa, outbreaks of African horse sickness (AHS) occur in summer; no cases are reported in winter, from July to September. The AHS virus (AHSV) is transmitted almost exclusively by Culicoides midges (Diptera: Ceratopogonidae), of which Culicoides imicola is considered to be the most important vector. The over-wintering mechanism of AHSV is unknown. In this study, more than 500 000 Culicoides midges belonging to at least 26 species were collected in 88 light traps at weekly intervals between July 2010 and September 2011 near horses in the Onderstepoort area of South Africa. The dominant species was C. imicola. Despite relatively low temperatures and frost, at least 17 species, including C. imicola, were collected throughout winter (June–August). Although the mean number of midges per night fell from > 50 000 (March) to < 100 (July and August), no midge-free periods were found. This study,using virus isolation on cell cultures and a reverse transcriptase polymerase chain reaction (RT-PCR) assay, confirmed low infection prevalence in field midges and that the detection of virus correlated to high numbers. Although no virus was detected during this winter period,continuous adult activity indicated that transmission can potentially occur. The absence of AHSV in the midges during winter can be ascribed to the relatively low numbers collected coupled to low infection prevalence, low virus replication rates and low virus titres in the potentially infected midges. Cases of AHS in susceptible animals are likely to start as soon as Culicoides populations reach a critical level

The virtual absence of Culicoides imicola (Diptera: Ceratopogonidae) in a light-trap survey of the colder, high-lying area of the eastern Orange Free State, South Africa, and implications for the transmission of arboviruses

Altogether 52 078 Culicoides biting midges of 35 species were collected during February 1990 and 1993 in 40 light-trap collections made on 17 cattle and/or sheep farms in the Bethlehem and Fouriesburg districts of the colder, high-lying eastern Orange Free State. Culicoides (Avaritia) bolitinos was by far the most abundant species, representing 50,9% of all specimens collected. Culicoides (A.) imicola, considered to be the most common stock-associated species in the summer rainfall areas of southern Africa, and the only proven vector of bluetongue virus (BTV) and African horsesickness virus (AHSV) in the subregion, was uncommon, comprising only 1,4%. While AHS is apparently absent, BT and bovine ephemeral fever (BEF) are endemic in this cooler, high-lying area of South Africa. The virtual absence of C. imicola implies that other Culicoides species, such as C. bolitinos and C. cornutus, may be involved in transmitting BT virus (and perhaps BEF) in the eastern Orange Free State, and possibly elsewhere in Africa. Virus isolation attempts made on 45 single species pools of C. bolitinos, C. pycnostictus, C. milnei, C. leucostictus, C. zuluensis and C. gulbenkiani were, however, negative. Finally, 20 of 28 blood-engorged Culicoides of 11 species, which were tested against cattle, sheep, horse, pig and bird antisera, tested only positive against cattle antisera.The articles have been scanned in colour with a HP Scanjet 5590; 600dpi. Adobe Acrobat XI Pro was used to OCR the text and also for the merging and conversion to the final presentation PDF-format.mn201

Possible over-wintering of bluetongue virus in Culicoides populations in the Onderstepoort area, Gauteng, South Africa

Several studies have demonstrated the ability of certain viruses to overwinter in arthropod vectors. The over-wintering mechanism of bluetongue virus (BTV) is unknown. One hypothesis is over-wintering within adult Culicoides midges (Diptera; Ceratopogonidae) that survive mild winters where temperatures seldom drop below 10 °C. The reduced activity of midges and the absence of outbreaks during winter may create the impression that the virus has disappeared from an area. Light traps were used in close association with horses to collect Culicoides midges from July 2010 to September 2011 in the Onderstepoort area, in Gauteng Province, South Africa. More than 500 000 Culicoides midges were collected from 88 collections and sorted to species level, revealing 26 different Culicoides species. Culicoides midges were present throughout the 15 month study. Nine Culicoides species potentially capable of transmitting BTV were present during the winter months. Midges were screened for the presence of BTV ribonucleic acid (RNA) with the aid of a real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) assay. In total 91.2% of midge pools tested positive for BTV RNA. PCR results were compared with previous virus isolation results (VI) that demonstrated the presence of viruses in summer and autumn months. The results indicate that BTV-infected Culicoides vectors are present throughout the year in the study area. Viral RNA-positive midges were also found throughout the year with VI positive midge pools only in summer and early autumn. Midges that survive mild winter temperatures could therefore harbour BTV but with a decreased vector capacity. When the population size, biting rate and viral replication decrease, it could stop BTV transmission. Over-wintering of BTV in the Onderstepoort region could therefore result in re-emergence because of increased vector activity rather than reintroduction from outside the region

Seasonal abundance and parity of Culicoides biting midges associated with livestock at Roma, Lesotho (Diptera : Ceratopogonidae)

A light-trap survey was undertaken of the species composition, seasonal abundance and parity of Culicoides at Roma, Lesotho, to establish whether the likely vectors for bluetongue and African horse sickness occur in this area as well as the chance of transmission. A total of 34 catches was made between 21 September 1985 and 24 September 1986; 32 819 Culicoides were caught belonging to 19 species. Culicoides numbers rapidly built up from December to a peak in February which implies that this may also be the optimum time for virus transmission. The number of Culicoides dropped sharply in April with the onset of cooler conditions. C. zuluensis was the dominant species forming 69,6 % of the totalled catches, followed by C. pycnostictus with 11,7%. C. imicola, the only proven vector of bluetongue, was never abundant representing only 4,4% of the midges caught. The parous rate for each of the 2 commonest species was low, implying a low vector capacity.The articles have been scanned in colour with a HP Scanjet 5590; 600dpi. Adobe Acrobat XI Pro was used to OCR the text and also for the merging and conversion to the final presentation PDF-format.lmchunu2014mn2014mn201

Use of real-time quantitative reverse transcription polymerase chain reaction for the detection of African horse sickness virus replication in Culicoides imicola

Despite its important role as vector for African horse sickness virus (AHSV), very little information is available on the dissemination of this virus in Culicoides (Avaritia) imicola Kieffer (Diptera: Ceratopogonidae). This study reports on the applicability of a real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) to detect AHSV in dissected midges. A total of 96 midges were fed on AHSV-infected blood, after which one test group was dissected into head/thorax and abdomen segments immediately after feeding and the other only after 10 days of incubation. The majority of the midges (96%) ingested the virus successfully and there was no significant difference between the virus concentration in the heads/thoraxes and the abdomens immediately after feeding. After incubation, virus was detected in 51% of the midges and it was confined to the abdomen in the majority of these. The fact that virus was detected only in the heads/thoraxes of four Culicoides midges after incubation suggests the presence of a mesenteronal escape barrier. Replication in the salivary glands was not shown. An increase of the mean virus concentration in the abdomen after incubation indicates localised viral replication. The real-time RT-qPCR is recommended for further studies investigating the replication and dissemination of AHSV in Culicoides midges