Widespread Circulation of Flaviviruses in Horses and Birds in Northeastern Spain (Catalonia) between 2010 and 2019.

The surveillance for West Nile virus (WNV) in Catalonia (northeastern Spain) has consistently detected flaviviruses not identified as WNV. With the aim of characterizing the flaviviruses circulating in Catalonia, serum samples from birds and horses collected between 2010 and 2019 and positive by panflavivirus competition ELISA (cELISA) were analyzed by microneutralization test (MNT) against different flaviviruses. A third of the samples tested were inconclusive by MNT, highlighting the limitations of current diagnostic techniques. Our results evidenced the widespread circulation of flaviviruses, in particular WNV, but also Usutu virus (USUV), and suggest that chicken and horses could serve as sentinels for both viruses. In several regions, WNV and USUV overlapped, but no significant geographical aggregation was observed. Bagaza virus (BAGV) was not detected in birds, while positivity to tick-borne encephalitis virus (TBEV) was sporadically detected in horses although no endemic foci were observed. So far, no human infections by WNV, USUV, or TBEV have been reported in Catalonia. However, these zoonotic flaviviruses need to be kept under surveillance, ideally within a One Health framework.

Mycobacterium microti Infection in Free-Ranging Wild Boar, Spain, 2017-2019.

Mycobacterium microti is a member of the Mycobacterium tuberculosis complex that causes pathology in many mammals. M. microti infections have been found in some countries in Europe. We report an outbreak of tuberculosis caused by M. microti in wild boars in Spain.

Usefulness of Eurasian Magpies (Pica pica) for West Nile virus Surveillance in Non-Endemic and Endemic Situations.

: In September 2017, passive surveillance allowed the detection of West Nile virus (WNV) lineage 2 for the first time in northern Spain in a northern goshawk (Accipiter gentilis). However, a cross sectional study carried out in Eurasian magpies (Pica pica) in a nearby area evidenced that WNV had been circulating two months earlier. Therefore, active surveillance in Eurasian magpies proved its effectiveness for the early detection of WNV in a non-endemic area. Further surveys in 2018 and the beginning of 2019 using young magpies (i.e., born after 2017) showed the repeated circulation of WNV in the same region in the following transmission season. Therefore, active surveillance in Eurasian magpies as well proved to be useful for the detection of WNV circulation in areas that may be considered as endemic. In this manuscript we present the results of those studies and discuss reasons that make the Eurasian magpies an ideal species for the surveillance of WNV, both in endemic and non-endemic areas.

Detection of West Nile virus lineage 2 in North-Eastern Spain (Catalonia).

In September 2017, West Nile virus (WNV) lineage 2 was detected in Catalonia (Northern Spain) in northern goshawks by passive surveillance. The phylogenetic analyses showed that it was related to the Central/Southern European strains, evidencing WNV lineage 2 spread to Western Europe. WNV local transmission was later detected in bearded vultures housed at the Wildlife Recovery center where the goshawk was transferred to. Further studies, before the following period of high mosquito activity, indicated that WNV had circulated intensively in poultry and horses but only surrounding of the area where the virus was detected. In other areas of Catalonia, circulation of flaviviruses different to WNV was identified. Public Health investigations failed to detect WNV infection in humans.

Six-year surveillance of Newcastle disease virus in wild birds in north-eastern Spain (Catalonia).

Given that Newcastle disease (ND) is one of the major threats for the poultry industry, testing of Newcastle disease virus (NDV) has been carried out since 2010 in cases of mortality in wild birds (passive surveillance) in Catalonia. The objective is to provide an early warning system to prevent the infection of poultry. Since 2010, 35 episodes of mortality in wild birds were attributed to NDV infection. Throughout this period there was a progressive expansion of NDV to new areas, with an increase in the episodes of mortality, although it is not clear whether they were the result of the spread of the virus, or of the improvement of the surveillance. Phylogenetic analyses indicate that two distinct sublineages of NDV, 4a and 4b, were circulating in Catalonia. Both sublineages seem to be endemic in the wild bird population, affecting mainly Eurasian-collared doves, with a clear pattern in relation to its spatial distribution (coincident with the distribution of this species), and its temporal distribution (with the majority of cases between September and February). So far, endemicity in wild birds has not resulted in ND outbreaks in poultry. However, there are still many uncertainties about, for example, whether NDV may expand to new areas of Catalonia (with higher poultry density), or about the threat that the apparently more novel sublineage 4a may represent. Hence, efforts should be made so that measures to prevent infection of poultry farms (particularly in high-risk areas and periods) are encouraged, and surveillance is maintained.

Intestinal motor disorders associated with cyclical bacterial overgrowth in a rat model of enteritis.

The aims of this study were: 1) to obtain an experimental model reproducing the characteristics of chronicity and spontaneous relapses found in inflammatory bowel disease (IBD) and 2) to correlate these changes with intestinal motility and bacteria translocation. For this purpose, two groups of Sprague-Dawley rats were used: a treated group that received two subcutaneous injections of indomethacin (7.5 mg/kg) 48 h apart and a control group that received saline. Blood leukocytes, TNF, and fecal parameters were monitored for 90 days after treatment. In treated rats, a cyclic oscillation of blood leukocytes and TNF concomitant with an inverse correlation of fecal output was observed. Treated rats were then selected either during their highest or lowest blood leukocyte values for motor activity and microbiological evaluation. Controls were obtained in age-matched rats. Rats with high leukocyte levels showed a decrease of motor activity. In contrast, animals with low leukocyte levels presented hypermotility. Bacterial overgrowth accompanied by bacterial translocation was found in the group with high leukocytes, whereas no differences were observed between the control and indomethacin groups during the lowest leukocyte phase. We obtained a model of IBD characterized by a chronic cyclic oscillation of intestinal motility, flora, and inflammatory blood parameters. During the high-leukocyte stage, motor activity decrease is related to bacterial translocation. This phase is followed by a reactive one characterized by hypermotility associated with a decrease in both bacterial growth and leukocytes. However, as in IBD, this reaction seems unable to prevent a return to relapse.

Porcine reproductive and respiratory syndrome virus (PRRSV) infection status in pigs naturally affected with post-weaning multisystemic wasting syndrome (PMWS) in Spain.

The purpose of the study reported here was to determine the prevalence of porcine reproductive and respiratory syndrome virus (PRRSV) in pigs affected with post-weaning multisystemic wasting syndrome (PMWS), a disease believed to be caused by porcine circovirus type 2 (PCV2). From May 1997 to February 2000, PMWS was diagnosed in 277 pigs (from 120 farms) submitted to the Veterinary Pathology Diagnostic Service, Veterinary School of Barcelona, Spain. In each case, the PMWS diagnosis was based on clinical history and the detection, by in situ hybridization, of nucleic acid of PCV2 in characteristic histologic lesions. Antigens for PRRSV were detected by immunohistochemistry in tissues of 66 (23.8%) of the same 277 pigs. Sera, which were available for 93 of the 277 pigs, were tested for PRRSV by a multiplex reverse transcription-polymerase chain reaction (RT-PCR). A total of 33 of these sera were RT-PCR positive, three for a North American strain(s) of PRRSV. In addition, 76 of the 93 sera were tested for antibodies to PCV2 (indirect immunoperoxidase) and PRRSV (enzyme-linked immunoassay). Antibodies for PCV2 and PRRSV were detected, respectively, in 56 (73.9%) and 43 (56.6%) of the 76 sera. Collectively, these results suggest that while infection with PRRSV may be common, it is not an essential component of PMWS.