Filogenetska i molekularna analiza gena hemaglutinina virusa ptičje gripe h9n2 izoliranih u jatima peradi u Maroku između 2016. i 2018. godine

Avian influenza viruses of the H9N2 subtype continue to spread in wild birds and poultry worldwide. Infection with H9N2 avian influenza virus was detected for the first time in Morocco in January 2016. In this study, a total of 105 organ and tracheal swab samples from 21 broiler farms in Morocco were collected from July 2016 to October 2018 for H9N2 screening. The suspicion of disease was based on severe respiratory signs such as sneezing, coughing, rales and gasping, while H9N2 virus infection was confirmed by real-time RT-PCR. Hemagglutinin (HA) genes of four isolates were amplified by conventional RT-PCR, sequenced, and aligned for phylogenetic analyses. Among the 21 flocks, 48% (10/21) were qRT-PCR positive for H9, with the cycle threshold values ranging from18.6 to 34.8. The maximum similarity in nucleotide and protein sequences (96-98%) was observed between the Moroccan viruses and an H9 virus isolated from broiler chickens in 2017 in Burkina Faso (A/chicken/BurkinaFaso/17RS93-19/2017) and from a layer chicken in the United Arab Emirates in 2015 (A/chicken/Dubai/D2506/2015). The HA genes revealed the close relationship between the four Moroccan viruses, with 97.9%-99.9% nucleotide identity. Phylogenetic analysis showed that the Moroccan viruses belonged to the G1 lineage, and likely originated from the Middle East, as previously reported in 2016.Virusi ptičje gripe H9N2 nastavljaju se širiti u peradi i divljih ptica širom svijeta. Infekcija niskopatogenim virusom influence H9N2 prvi je put otkrivena u Maroku u siječnju 2016. godine. U ovom je istraživanju za probir na H9N2 prikupljeno ukupno 105 organa i obrisaka iz dušnika s 21 farme brojlera od srpnja 2016. do listopada 2018. iz različitih regija Maroka. Sumnja na bolest temeljila se na teškim respiracijskim znakovima kao što su kihanje, kašljanje, hropanje i hripanje, a infekcija virusom H9N2 potvrđena je PCR-om obrnute transkripcije u stvarnom vremenu. Sekvencije gena za hemaglutinin (HA) od četiri izolata amplificirane su pomoću RT-PCR qRT-PCR poravnane za filogenetsku i analizu sličnosti aminokiselina. Od 21 uzorka jata 48 % (10/21) bilo je pozitivno na H9 s pragom broja ciklusa u rasponu od 18,6 do 34,8. Maksimalna sličnost u nukleotidnim i proteinskim sekvencijama (96 -98 %) uočena je između marokanskih virusa i virusa H9 izoliranih iz brojlerskih pilića u 2017. u Burkini Faso (A/piletina/BurkinaFaso/17RS93-19) i od kokošjeg pileta u Ujedinjenim Arapskim Emiratima u 2015. (A/piletina/ Dubai/D2506/2015). HA geni otkrili su blisku vezu između četiriju virusa, s 97,9 % -99,9 % nukleotidnog identiteta. Filogenetska analiza pokazala je da marokanski virusi pripadaju lozi G1 i vjerojatno potječu s Bliskog istoka, kao što je objavljeno 2016. godine

A Century Spent Combating Rabies in Morocco (1911–2015): How Much Longer?

Rabies has no known beginning in Morocco and to date, government control efforts and plans fail to eradicate the disease. A review and analysis of available epidemiological data are crucial to learn lessons from the past and to propose effective actions. Legally, animal rabies is a notifiable disease since 1913 and legislation has been updated periodically since. Dogs have always been considered as both the disease’s vector and reservoir, while cattle, other herbivores, and humans are victims. Animal rabies cases evolution from 1942 to 2015 is characterized by ascending phase then decreasing one following structured rabies control plan implementation in 1980s. Indeed, from 1986 to 2010, three rabies control plans have been conducted based on free of charge rabies vaccination of owned dogs through mass campaigns. The geographical distribution of rabies is stable over the years with highest cases number in rich rural areas and around cities. Human rabies cases are decreasing over the time (1976–2015) thanks to the opening of new antirabic treatment centers in the last decade which permit the administration of more PEPs. After a century of rabies control, Morocco registered an average of 301 animal cases and 21 human cases annually for the last decade (2005–2015). Few reasons led to those limited results. The lack in law enforcement and, moreover, the fact that the law do not take into account responsible dog ownership aspect are of importance. Lack of dog population knowledge and management and intersectoral coordination deficiency are additional failure reasons. The gathered data will help to build a new strategy with a focus on a “One Health” approach. Dog population ecology parameters’ study is of primary importance. We estimated dog population to be 2.8 million dogs based on human:dog ratio. Enhancing vaccination coverage of dog population is feasible by combining parenteral vaccination and complementary oral vaccination. Updating legislation by inclusion of responsible dog ownership and law enforcement are crucial. Over the last century, Morocco registered a slow decreasing tendency in the number of animal and human rabies cases. Urgent strategy need to be implemented because rabies elimination is an achievable goal in Morocco

Evaluation of Bait Acceptance and Immune Response in Local Dogs during an Oral Rabies Vaccination Field Study in Morocco

The objective of this study was to evaluate the bait preference of three selected bait types by local dogs and the induced immunogenicity of the oral rabies vaccine strain SPBN GASGAS in Morocco. The vaccine strain, combined with different bait types, has been tested in many different settings, but not yet in northern Africa. Overall, bait consumption and preference were similar in other studies using the same materials (bait type and sachet). The intestine bait had the highest acceptance rate (97.6%, 95%CI: 87.4–99.9), followed by the egg bait (83.0%, 95%CI: 69.2–92.4). Only 52% (95%CI: 37.4–66.3) of the dogs showed an interest in the fish meal bait. However, considering the successful release of the contents of the sachet (blue-dyed water) into the oral cavity, the egg bait (65.7%, 95%CI: 47.8–80.9) scored better than the intestine bait (51.7%, 95%CI: 32.5–70.6). The dogs selected for the immunogenicity study were offered the egg bait containing a sachet filled with SPBN GASGAS (3.0 mL, 107.5 FFU/mL) or were given the same dose by direct oral administration (d.o.a.). In addition, several dogs were vaccinated by the parenteral route (s.c.) using a commercially available inactivated rabies vaccine. Unfortunately, due to the COVID-19 pandemic and subsequent travel restrictions, it was not possible to collect blood samples directly after vaccination. The blood samples were collected pre-vaccination and on five occasions between 450 and 1088 days post vaccination. The seroconversion rate, as determined for rabies-virus-neutralizing antibodies by the FAVN test, was significantly lower than that found for binding antibodies, as determined by ELISA, for all blood samples collected post vaccination. No treatment effect (bait, d.o.a., s.c.) could be seen in the seroconversion rate. At 15 months post vaccination, 84.2% of the dogs offered vaccine bait still tested sero-positive in ELISA. Only after 3 years was a clear drop in the seroconversion rate observed in all three treatment groups. This study confirms the long-term immunogenicity of the oral rabies vaccine SPBN GASGAS in dogs under field conditions