Urgent request on avian influenza.

Highly pathogenic avian influenza (HPAI) H5N8 is currently causing an epizootic in Europe, infecting many poultry holdings as well as captive and wild bird species in more than 10 countries. Given the clear clinical manifestation, passive surveillance is considered the most effective means of detecting infected wild and domestic birds. Testing samples from new species and non-previously reported areas is key to determine the geographic spread of HPAIV H5N8 2016 in wild birds. Testing limited numbers of dead wild birds in previously reported areas is useful when it is relevant to know whether the virus is still present in the area or not, e.g. before restrictive measures in poultry are to be lifted. To prevent introduction of HPAIV from wild birds into poultry, strict biosecurity implemented and maintained by the poultry farmers is the most important measure. Providing holding-specific biosecurity guidance is strongly recommended as it is expected to have a high impact on the achieved biosecurity level of the holding. This is preferably done during peace time to increase preparedness for future outbreaks. The location and size of control and in particular monitoring areas for poultry associated with positive wild bird findings are best based on knowledge of the wider habitat and flight distance of the affected wild bird species. It is recommended to increase awareness among poultry farmers in these established areas in order to enhance passive surveillance and to implement enhanced biosecurity measures including poultry confinement. There is no scientific evidence suggesting a different effectiveness of the protection measures on the introduction into poultry holdings and subsequent spread of HPAIV when applied to H5N8, H5N1 or other notifiable HPAI viruses.

Epidemiological aspects of viral haemorrhagic septicaemia virus genotype II isolated from Baltic herring, Clupea harengus membras L.

This study was carried out to clarify the role of wild fish, especially Baltic herring, Clupea harengus membras L., in the epidemiology of viral haemorrhagic septicaemia virus (VHSV) in brackish water in Finland. Baltic herring with no visible signs of disease were collected from the Archipelago Sea, the Gulf of Bothnia and the eastern Gulf of Finland. In total, 7580 herring were examined by virus isolation as 758 pooled samples and 3029 wild salmonid broodfish as pooled samples during 2004-2006. VHSV was isolated from 51 pooled herring samples in bluegill fibroblast-2 cells, but not in epithelioma papulosum cyprini cells. The majority of isolations were from the coastal archipelago and from fish caught during the spawning season. Based on glycoprotein (G) gene sequences, the virus was classified as a member of genotype II of VHSV. Pairwise comparisons of the G gene regions of herring isolates revealed that all the isolates were closely related, with 98.8-100% nucleotide homology. Phylogenetic analyses revealed that they were closely related to the strains isolated previously from herring and sprat, Sprattus sprattus (L.), in Gotland and to the VHSV isolates from European river lamprey, Lampetra fluviatilis (L.), in the rivers that flow into the Bothnian Bay. The infection in Baltic herring is likely to be independent of the VHSV Id epidemic in farmed rainbow trout, Oncorhynchus mykiss (Walbaum).

Yersinia enterocolitica and Y. enterocolitica-like species in clinical stool specimens of humans: identification and prevalence of bio/serotypes in Finland.

This study investigated the prevalence of Yersinia enterocolitica (YE) bio/serotypes and YE-like species in clinical stool specimens. The special aim was to find the best methods for accurate identification of YE species and, further, pathogenic strains among YE isolates. Of the 41,848 specimens cultured in ten laboratories during a 12-month period, 473 Yersinia strains were isolated from 462 patients. The strains were identified by 21 biochemical tests, serotyping, colony morphology, as well as by 16S rRNA and gyrB gene sequencing. The most prevalent Yersinia findings were YE biotype 1A (64% of the strains) and pathogenic bio/serotype 4/O:3 (16%). The cold-enrichment increased the number of all isolates, and 25% of the bio/serotype 4/O:3 and 2/O:9 strains were only found by cold-enrichment. In routine diagnostic laboratories, 50% of the YE-like species were identified as YE and in 26% the identification differed from that of the reference laboratory. The microscopic colony identification on CIN agar with positive CR-MOX test, combined with several biochemical tests, identified reliably the pathogenic YE bioserotypes and most YE BT 1A strains, but some strains of the YE-like species were so heterogenic that gene sequencing was the only way to identify them.

Transmission of bovine viral diarrhoea virus through the semen of acutely infected bulls under field conditions.

Epidemiological investigations implicated the semen of artificial insemination (ai) bulls as the only plausible source of infection with bovine viral diarrhoea virus (bvdv) in 10 Finnish dairy herds. The infection was traced back to two northern Finncattle bulls that had been transiently infected when their semen had been collected while they were in a gene bank herd containing persistently infected (pi) animals. The isolates of bvdv from the animals in the gene bank herd, from the semen of the two bulls and from a pi calf born in one of the herds using the semen belonged to a rare genetic type in Finland and, on the basis of the nucleotide sequences in the 5' untranslated region, were identical. Cross-contamination of batches of semen at the ai station and an external source of bvdv were ruled out for the recipient herds. It was concluded that bvdv infection can be transmitted through the semen of transiently infected bulls under field conditions.

Etiology of respiratory disease in non-vaccinated, non-medicated calves in rearing herds.

The aim of this study was to examine the occurrence of bacterial, mycoplasmal and viral pathogens in the lower respiratory tract of calves in all-in all-out calf-rearing units. According to clinical status, non-medicated calves with and without respiratory disease signs were selected of the 40 herds investigated to analyse the micro-organisms present in healthy and diseased calves. Tracheobronchial lavage (TBL) and paired serum samples were analysed for bacteria, mycoplasmas, respiratory syncytial virus (RSV), parainfluenza virus 3 (PIV3), bovine corona virus (BCV) and bovine adenovirus (BAV). Pasteurella multocida was the most common bacterial pathogen. It was isolated from 34% of the TBL samples in 28 herds and was associated with clinical respiratory disease (p < 0.05) when other pathogenic bacteria or mycoplasma were present in the sample. Mannheimia spp. and Histophilus somni were rarely found. Mycoplasma bovis was not detected at all. Ureaplasma diversum was associated with clinical respiratory disease (p < 0.05). TBL samples from healthy or suspect calves were more often negative in bacterial culture than samples from diseased calves (p < 0.05). No viral infections were detected in six herds, while 16-21 herds had RSV, BCV, BAV or PIV3. In the herds that had calves seroconverted to BCV, respiratory shedding of BCV was more frequently observed than faecal shedding. This study showed that the microbial combinations behind BRD were diverse between herds. M. bovis, an emerging pathogen in many countries, was not detected.

Eradication of enzootic bovine leukosis from Finland.

Enzootic bovine leukosis (EBL) was recognized among Finnish cattle in 1966. Administrative decisions specifying and refining official control measures were given in 1966, 1976, 1980, and 1993. The measures' key principle always has been 'test and slaughter'. The EBL/bovine leukosis virus (BLV) infection situation was monitored at meat inspection, and hematologically between 1970 and 1977 and serologically between 1978 and 1989. Annual surveys including all dairy herds and samples from beef animals were conducted in 1990-2001. Bulk-tank milk samples represented the dairy herds in the surveys; the beef animals were sampled individually at slaughter. The maximum positive herd-level percentage in the surveys was 0.03%. EBL/BLV infection was evenly dispersed in the southern part of the country and nonexistent in the northern part. We conclude that herd-level prevalence of EBL/BLV infection never exceeded 5%. It nevertheless took 30 years to eradicate the disease and the infection. EBL was eradicated from mainland Finland in 1996 and from the island district of Ahvenanmaa in 1999. Annual monitoring of the EBL situation continues.

Distemper vaccination of farmed fur animals in Finland.

The most important farmed fur animal species in Finland are the American mink (Mustela vison), blue fox (Alopex lagopus), silver fox (Vulpes vulpes) and raccoon dog (Nyctereutes procyonoides); all are susceptible to canine distemper. The only distemper vaccines currently available are for mink, although they also have been used for fox and raccoon dogs in emergency situations. The efficacy in eliciting neutralizing antibodies and the safety of three mink-distemper vaccines were studied under field conditions with mink and silver fox. Two of the vaccines were also studied with raccoon dogs and blue fox. All three vaccines elicited a satisfactory antibody response in mink, whereas the response varied in the other species. No side effects were observed in any species tested. One of the vaccines was safe and immunogenic in all four species.

Documenting freedom from disease and re-establishing a free status after a breakdown. Rabies.

Rabies reappeared in Finland in the spring of 1988 after a 29-year absence. This time rabies occurred in sylvatic form and the major species involved was the raccoon dog. During the outbreak 1988-89 66 animals were diagnosed rabid. Vaccination of cats, cattle and horses was strongly recommended and vaccination of dogs was compulsory in the outbreak area. A field trial was started on oral immunisation of raccoon dogs and foxes against rabies using baits containing rabies vaccine strain. The outbreak area and a wide buffer zone were baited three times. Finland was declared free of rabies again in 1991. Oral vaccination campaign with vaccine baits has been organised along the south-eastern border once a year since the beginning of 90s. Continuous surveillance and epidemiological screening is necessary to detect any new outbreaks of rabies at an early stage.

Canine distemper virus neutralising antibodies in vaccinated dogs.

The associations between the levels of canine distemper virus neutralising antibodies and vaccination history, age and gender were investigated in a cross-sectional study of a sample of 4627 dogs from the Finnish urban dog population. Dogs vaccinated with either Canlan (Langford Laboratories) or Dohyvac (Solvay Animal Health) or with both, had significantly lower titres than those vaccinated with Candur (Behringwerke), Duramune (Fort Dodge Laboratories) or Nobivac (Intervet), or with combinations including at least one of these. The vaccines were classified as having low and high immunogenicity on the basis of the geometric mean titre achieved by the vaccine when compared with the geometric mean titre of the entire dataset. The low geometric mean titre of Canlan, Dohyvac and their combination groups resulted from the large proportion of dogs without detectable titres, especially dogs under one year of age, rather than from uniformly low titres. An age-stratified comparison of vaccine usage and titres showed that the division of the vaccines into low and high immunogenicity vaccines was apparent in the dogs less than two years of age but not in the older dogs. The first vaccination with the high immunogenicity vaccines resulted in a higher proportion of dogs with detectable antibodies than even repeated vaccination with the low immunogenicity vaccines. Neither the time elapsed since the most recent vaccination nor the gender of the dogs was associated with the titre of antibody.

Preventing the spread of maedi-visna in sheep through a voluntary control programme in Finland.

The sheep disease maedi-visna (MV) was introduced into Finland in 1981 and had spread to eight flocks in the southwestern part of the country when first detected in a survey in 1994. Six more seropositive flocks were subsequently traced, bringing the total to 14. MV has a notifiable disease status in Finland that provides for official restrictive measures to which all infected herds are subject. These measures are withdrawn once the seropositive animals and their progeny are culled and the flock has showed negative signs in the test done twice, or after total culling. A voluntary control programme was initiated in January 1995 to extend official control efforts. The programme furnishes a guideline for culling, restrictions on contacts, and a timetable for testing the flock to attain MV-free status. Seven flocks of the 14 were slaughtered either immediately or after a period under restrictive measures. One flock finished sheep production after four years under restrictive measures. Selective culling and repeated testing was attempted with the other six flocks, three of which attained MV-free status. One flock finished sheep production after two years in the control programme, the other two dropped out of the programme when the restrictive measures were withdrawn. It was concluded that the control programme was salient in eradicating MV from Finland and that serological monitoring of the situation must be continuous.

Serological survey and epidemiological investigation of maedi-visna in sheep in Finland.

A survey for antibodies to maedi-visna virus (MV) in the Finnish sheep surveillance flocks was conducted in 1994. Examination of a total of 12931 serum samples from animals over 1 year of age from 545 flocks (81% of all flocks) revealed eight seropositive flocks and the subsequent epidemiological investigation yielded one additional seropositive flock, indicating a low prevalence of 1.6%. The infection was very probably imported from Sweden in 1981, but it was not detected until the survey was conducted 13 years later. The entire primary infection flock was slaughtered in 1995. 77% of the sheep were seropositive but the animals were clinically healthy and only one (5%) of the contact flocks of the primary infection flock had contracted the infection. This secondary infection flock, 77% of which was seropositive, was slaughtered in 1994; however, animals in this flock had respiratory problems and the lungs of three sheep showed typical MV lesions. Seven (24%) of its contact flocks had contracted the infection and these each had one or two seropositive animals except for one flock which had seven (18%) seropositive animals. The results show that the initial spread of MV can be insidious and wide before infection is revealed in surveys or any clinical cases are encountered.

Prevalence and geographic distribution of bovine viral diarrhoea (BVD) infection in Finland 1993-1997.

Bulk milk samples from every herd supplying milk to dairies in Finland were examined for the presence of antibodies to BVD virus (BVDV) annually during 1993-1997. The highest prevalence, 0.99% in 1994, declined to 0.37% in 1996; however, this favourable trend appeared to discontinue in 1997, where the prevalence remained at 0.41%. In 1993, sera of all individual animals from bulk milk antibody-positive herds were examined for the presence of these antibodies. Since 1994, only sera of animals from herds with a bulk milk absorbance reading greater than 0.250 in the EIA test were examined individually. Three geographic foci of BVDV antibody-positive dairy herds were resolved in 1994, one in the north-western, another in the eastern and a diffuse third in the southern part of Finland. A distinct limiting of the spread was apparent in 1997. Beef cattle were also studied during 1993-1997; in 1993 breeding units, in 1994 mainly beef suckler herds and in 1995-1997 serum samples of beef animals at slaughter were examined for the presence of antibodies to BVDV. The prevalence of seropositive herds in 1993 and 1994 was 30.2% and 3.2%, respectively, while the prevalence among slaughter animals ranged 0.8-1.6%. Seronegative animals in herds with > 50% of seropositive animals were examined for the presence of BVD-virus. A total of 40 dairy herds and two beef herds with viraemic (persistently infected, PI) animals was encountered during 1993-1997. A comprehensive control programme and a more specific, cooperatively funded eradication programme for dairy cattle were launched in 1994. These programmes most probably contributed to the decline in prevalence during 1994-1996.

Outbreak off canine distemper in vaccinated dogs in Finland.

Canine distemper reappeared in dogs in Finland in 1990 after a 16-year absence. In 1994 to 1995 an outbreak occurred in areas with a high density dog population which involved dogs vaccinated against distemper. The estimated total number of cases was at least 5000, and 865 cases were confirmed by indirect fluorescent antibody testing of 3649 epithelial cell samples. The signs recorded by veterinary clinicians ranged from conjunctivitis, pyrexia and anorexia to signs of respiratory and gastrointestinal illness, with an estimated mortality of 30 per cent. Of the confirmed cases 631 (73 per cent) were between three and 24 months of age; 487 of these had been vaccinated at least once and 351 (41 per cent) had a complete vaccination history. Of these 351 fully vaccinated animals the proportion of dogs vaccinated with the most popular vaccine was significantly higher than would have been expected by its market share. In total, 4676 serum samples were collected from healthy vaccinated dogs during the peak and decline of the outbreak and tested for the presence of virus neutralising antibodies. The decrease in the proportion of young dogs with antibody titres < 1/8 coincided with the decline and end of the outbreak during the spring and summer of 1995. It was concluded that a critical decrease in the population's immunity during 1990 to 1994 was a major reason for the outbreak in the summer of 1994 and that the ultimate test for vaccines is an outbreak of disease.

Rabies antibodies in vaccinated dogs.

Forty-seven healthy, owned dogs were vaccinated with Madivak and 85 with Rabisin. Geometric mean titres of 17.40 and 1.03 IU/ml were measured by the rapid immunofluorescent focus inhibition test 30-40 and 350-370 days, respectively, after a single injection. Four out of 130 (3.1%) and 18 out of 106 (17%) dogs had a titre of less than 0.5 IU/ml in serum 30-40 and 350-370 days after vaccination. Twenty-one dogs (19.8%) had a titre of 0.5 IU/ml 350-370 days after vaccination. There was no significant difference in antibody levels between animals vaccinated with Rabisin or Madivak. Our results indicate that a booster is always necessary after a single injection to ensure that all dogs have a lasting antibody titre.

Immunization of cattle against rabies using inactivated cell culture vaccines.

Twenty-one heads of cattle were vaccinated with Madibovin, 31 with Rabdomun and 127 with Rabisin on 4 different farms. Rabies neutralizing antibody titre (> or = 0.5 IU/ml) was detected in 80% of 163 animals tested about 1 month and in 42% of 133 animals tested about 1 year after primary vaccination. On 3 of the farms 86 animals received booster vaccination about 1 year after primary vaccination. All these animals had antibody titre (> or = 0.5 IU/ml) about 1 month after booster and antibody levels were higher than after the primary vaccination. Rabies antibody titres (> or = 0.5 IU/ml) were detected in 96% of 50 animals tested 1 year after the booster. No significant differences (p > 0.05) in antibody levels were detected between animals vaccinated with Madibovin or Rabisin (farm C) respectively with Rabisin or Rabdomun (farm D) at any collection time. Responses to rabies vaccines varied considerably between the farms. After primary vaccination of the animals on 2 farms with the same batch of Rabisin, the antibody levels clearly differed (p < 0.0001) between the farms. Our results indicate that booster is always necessary after primary vaccination to ensure that all animals are protected.

Rabies antibody titres in vaccinated reindeer.

One dose of inactivated, adjuvanted rabies vaccine of cell culture origin (Rabisin) induced good but short-duration immunity in close to 100% of the 50 semi-domesticated reindeer (Rangifer tarandus tarandus L.) vaccinated. Most of the animals (44) had rabies virus antibody titre > or = 1.5 IU/ml at 38 days after vaccination. Five animals had titre 0.5 IU. Antibody titres were not, however, present 1 year after primary vaccination in most animals. About 1 year (360-413 days) after primary vaccination, 22 of the 39 reindeer that could be sampled had rabies virus antibody titre < 0.5 IU/ml.

Effects of eating an uncooked vegetable diet for 1 week.

Biochemical and metabolic indicators were monitored in a group of volunteers suffering from a variety of chronic illnesses participating in a week's course on a special uncooked vegetable diet, known as "living food". Unmatched healthy controls ate the same diet cooked for 2 min in a microwave oven. After 1 week on the regimen, serum protein and urea contents decreased and alanine aminotransferase (ALAT) activity increased in both groups, although all within the normal range. Blood glucose increased in both groups to slightly above normal limits but total serum cholesterol dropped about 1 mmol/l from normal starting levels. Serum tocopherol and retinol increased only in the group eating the uncooked diet. In both groups urinary sodium dropped drastically without a significant change in potassium. Serum and urinary phenol and p-cresol diminished also significantly. It is concluded that this vegetable diet may be of some benefit in the short term but any longer-term use requires evaluation.