1 Dromedaries as possible reservoir of some infectious diseases: the case of Brucellosis…and other viruses! Giovanni Savini Abu Dhabi February 15th 2016
2 We are here
3 Animal Health Food Safety Animal Welfare Information Systems
4 Our international expertise
5 My presentation Brief Introduction to camelids and related problemsMost important infectious disease of camelids at the animal/human interface (Rabies, Camelpox, Brucellosis) Our experience at the IZSAM with camelids (BTV, RVF, WNV) and our future goals
6 COMMON ORIGIN 60 millions years agoEvolution
7 NEW WORLD CAMELS OLD WORLD CAMELSSensisbilità diversa, legata alla distribuzione geografica?
8 Economic impact New world camels are very popular in Europe and UScamelids (llama and alpaca) are present in the US Are camelids susceptible to the wide range of viruses affecting large and small ruminants? Importanza economica è crescente interesse in Europa allevamenti di camelidi
9 Economic impact Pathogenic Viral InfectionsRabies, Borna, Camelpox, Parapox, Papillomatosis, FMD, VS, PPR, RFV, BVD, BT, WND, EEE, Rota/Corona (Diarrhoea), BoHV-1, EHV, Flu A, Equine rhinovirus abortion Non-Pathogenic Viral Infections Rinderpest, AHS, Respiratory viruses, Retrovirus infections
10 Antibodies for many viral agents were detected
11 Antibodies for many viral agents were detectedPresenza di positività sierologica non è indice di suscettibilità
12 In general camelids are relatively free of diseasesSono animali resistenti
13 RABIES Source: Incubation DeathFeral Dogs. Transmission from alpaca to alpaca by bites has been described Incubation NWCs: 15 days-3 months OWCs: 3 weeks to 6 months Death 6 to 8 days after clinical signs Rabbia importante, trasmissione diretta, sintomo classico tentativo di sbadiglio
14 RABIES Clinical signs in OWCs:Raging Fury for 1 or 3 days (male dromedary) followed by paralytic stage (attempt to yawn) Silent Fury, generally seldom seen in camels During Raging Fury: restless, aggression, selfmutilations biting and snapping, hypersalivation in NWCs: Pica, aggressive form, attacks on people pen mates and offspring. (No spit). Rabbia importante, trasmissione diretta, sintomo classico tentativo di sbadiglio
15 Brucellosis OWCs: frequently infected with brucellosisNWCs: rare but described Zoonosis: 500,000 new cases annually, 30% of camel handlers and 2% of Laboratory workers Infection: contact with infected material, unpasteurized milk and dairy products, uncontrolled trade, airborne transmission
16 Brucellosis Infection is related to the presence of sheep and cattlePlacentas and aborted foetuses are the most infected tissues Milk excretion seems to be intermittent in camels Brucella is very resistent even if it prefers cool temperature it can survive also in hot desert environment.
17 Brucellosis B. melitensis and B. abortus the most serious and the most common B. melitensis is common in camels in ME and Africa (biovars 1, 2, 3 and 6) B. abortus in the former URSS Incubation period 60 dd
18 Brucellosis Abortions (1° pregnancy) Stillborn calves Reduced milkRetained placenta has been never described Orchitis Placenta different, ruminants cotyledonary, camel diffusa
19 Brucellosis Diagnosis in the field can be difficult because of other causes of abortions: Salmonellosis Trypanosomosis Campylobacter spp. infection Trichomonas foetus
20 Brucellosis Serological assays:Molecular and culturing are the preferred test methods whereas multiple serological tests are needed for identification of all reactors Serological assays: CFT (longer reaction) RBT SAT (BPAT ecc…) (IgM and IgG) FPA (promising) ELISA ELISA on milk is also possible (modified MRT)
21 Brucellosis There is a need for a standardization of assays!!!!Serology only can be confusing (false positive by Y. enterocolitica) The combination of multiple tests is CRUCIAL (real time and serological tests, 99% sensitivity) None of the serological tests are validated for camel (human) There is a need for a standardization of assays!!!!
22 Brucellosis Antibiotic treatments are long and difficult and with doubtful results Vaccination more doubts than certainties Brucella abortus S19 vaccines Brucella melitensis Rev1 No challenge after vaccination Best age for vaccination Reduced dose, immunity lasts 3-4 months
23 Bluetongue AHSV (1-8) BTV (1-27) EHDV (1-7)?Live camel exportation risk of transboundary disease EHDV (1-7)? AHSV (1-8)
24 BTV-26, direct transmission
25 New Serotypes BTV-28 BTV-29 Middle East South Africa
26 Incursioni del virus della BT nel bacino del MediterraneoTo summarise our molecular epidemiology studies (and ignoring the vaccine strains of BTV-2, 4, 9 and 16 that have been used in the Mediterranean region) we have evidence that BTV has entered Europe on 9 separate occasions since 1998, via four distinct routes, including new introductions each year except 2002. There appear to have been introductions from the east, (possibly via Turkey), of BTV-1, 9 and 16. BTV 4 in Greece was very closely related to strains from Cyprus, going back to 1969, suggesting that the virus has been circulating there almost unnoticed for many years, supported by the local ecosystem. In contrast BTV 2 and a distinct strain of BTV-4 appear to have entered Europe directly from North Africa . Then in 2006 a new and different strain of BTV-1 arrived from North Africa, and BTV-8 parachuted into northern Europe. 26
27 sedimentarisation Figure 1: Main transhumance routes (in red) of small ruminants and camels across Morocco
28 Serosurveillance in Mauritania58% of cattle positive for BTV by c-ELISA 68% of dromedaries positive for BTV by c-ELISA It is necessary to get reliable data 47 animals had BTV 26 neutralising Abs 99 animals had BTV 26 neutralising Abs
29 Surveillance in Mauritania20 pos 1/27 20 neg 1/24 5 pos 1/27 5 neg 1/24 LOW TITRES LOW TITRES rt RT-PCR detecting 1/27 serotypes
30 Surveillance in MauritaniaBTV-26, Kuwait 2010 One single isolate Used for animal work Very low titres in goats and sheep
31 Surveillance in MauritaniaBTV-26 infected samples from Mauritania were not recognized as BTV-26 bt serotype specific rt-RT-PCR Divergent BTV-26 circulating in Africa? Novel serotype in Africa NGS did not help (low titres) Small piece of genomic information Poor quality of samples
32 Surveillance in Mauritania……. however, considering the serological results and that... Some samples were positive by ELISA and by rt RT-PCR, negative by SN Recent infections at the slaughterhouse (animals gathered together up to 10 days) Direct transmission
33 Origin BTV-26
34 Evolution
35 BTV-4 2012: Sardinia BTV-4 SAD2012 BTV-1 SAD201210 5 BTV-1 SAD2012 BTV-4 SAD2012 86% of confirmed cases 13% of confirmed cases
36 BTV-1, BTV-2, BTV-4, BTV-16 and BTV-24BTV : Balkan Countries and Italy BTV-1, BTV-2, BTV-4, BTV-16 and BTV-24 BTV-4 APU2014
37 BTV-4 2014: Morocco and SpainBTV-2, BTV-3, BTV-4, BTV-16 16 4 2 3 16 4 2 3 Morocco/1-2014 Morocco/2-2014
38 X Potential evolution of BTV BTV-1 or BTV-8 Direct contact?BTV1/8 Vector Borne strain 1 10 5 1 10 5 X BTV-1 or BTV-8 Direct contact?
39 BTV-26 in Mauritania
40 Surveillance in MauritaniaIn the framework of scientific collaboration between CNERV of Nouakchott-Mauritania and IZSA&M of Teramo-Italy, 157 bovine blood samples and 119 blood samples from camels collected at the Nouakchott abattoir in March 2013, have been tested by molecular tests for different pathogens , considered to be likely present or at risk in the Country Network of mediterranean basin countries
41 Surveillance in MauritaniaPathogens & tests Dromedaries Bovines SERA AHSV => c-ELISA EHDV => c-ELISA RVFV => IgG, c-ELISA RVFV => qRT-PCR PPRV => c-ELISA WNV => c-ELISA WNV => SN BLOOD BTV => qRT-PCRs VI SERA RVF => IgG, c-ELISA RVF => qRT-PCR PPRV => c-ELISA PPRV => SN BLOOD BTV => qRT-PCRs VI Informazioni importanti su malattie importanti
42 Surveillance in MauritaniaVector transmitted bacterial and parasitic pathogens Cattle Positives (percentage) Tested Trypanozoon subgenus (T. brucei, T. evansi and T equiperdum)-Realtime PCR Theileria annulata-Real time PCR E. ruminantium-traditional PCR 119 114 (96%) Camels Trypanozoon subgenus-Real time PCR-Confirmed by Variable Surface Glycoprotein RoTat 1.2 PCR specific for T.evansi Theileria annulata-Realtime PCR 157 8 (5.10%)
43 Surveillance in MauritaniaVirological results Camelidae AHS c-Elisa EHDV c-Elisa WND c-Elisa WND SN PPR c-ELISA RVF IgG c-ELISA RVF RT-PCR Pos 5 63 146 41 72 Doubt 1 2 3 4 Neg 153 21 12 70 159 83 Tested 86 138 * Other flaviviruses? ** * Typing pending ** 24 samples were toxic for cells
44 PPRV Camelids are susceptible to the infectionThey could play an important role by disseminating the infection (to goats) Clinical signs may be different: fever, nasal discharge but also sudden death, diarrhoea, abortion Morbillivirus like distemper, vettore di malattia, no registered vaccine PPR like virus
45 RVFV Brief period of viraemia Abortion is the only sign of infectionNo data on calves No data on Bactrian camels Described (?) in Saudi Arabia in 2000 and in Kenya in 2006 but the virus was never isolated An attenuated vaccine exists (strain MP-22) for dromedaries. Good humoral response but challenge was not performed Sintomatologia simile a RVF ma mai isolato. Lancelot vaccino attenuato per cammelli no challenge test outbreak in 2000 in saudi Arabia camel importaion Zoonosis
46 Surveillance in MauritaniaVirological results Camelidae AHS c-Elisa EHDV c-Elisa WND c-Elisa WND SN PPR c-ELISA RVF IgG c-ELISA RVF RT-PCR Pos 5 63 146 41 72 Doubt 1 2 3 4 Neg 153 21 12 70 159 83 Tested 86 138 * Other flaviviruses? ** * Typing pending ** 24 samples were toxic for cells
47 Neutralizing antibodies for WNVWND Neutralizing antibodies for WNV Number of positive Malattia non descritta, sintomi cerebrali in america 5 10 20 40 80 160 320 Titre NWCs get infected with WNV but in general the role of camelids needs to be investigated
48 Surveillance in MauritaniaVirological results Camelidae AHS c-Elisa EHDV c-Elisa WND c-Elisa WND SN PPR c-ELISA RVF IgG c-ELISA RVF RT-PCR Pos 5 63 146 41 72 Doubt 1 2 3 4 Neg 153 21 12 70 159 83 Tested 86 138 * Other flaviviruses? ** * Typing pending ** 24 samples were toxic for cells
49 Surveillance in MoroccoOrbivirus Serosurveillance % Seropositive Year Virus 2003 2009 2010 2011 2012 BTV 11% 25,59% 34,93% 26,87% 59,71% AHSV 1,28% 1,07% 1,20% 0,00% EHDV WND 7% % 2014
50 Future Goals at IZSAM Role of camelids as BTV and EHDV reservoirRole of camelids in the biological cycle of flaviviruses (Zika?) Validation of serological assays for camelids
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52 Conclusion Pragmatic attitudes should be adopted to imagine and implement new strategies. In front of new challenges experience and conservative thoughts might not be the optimal solution
53 شكرا لاهتمامكم