R. C. Mainar-Jaime

Seroprevalence of Neospora caninum and abortion in dairy cows in northern Spain

The seroprevalence of Neospora caninum infection was estimated from a sample of 889 cattle from 43 dairy herds in three counties in the Asturias region of Spain. The true prevalence of infection was estimated to be 30-6 per cent (95 per cent confidence interval (ca) 27.6 to 33.6). Seropositivity was associated with abortion during the previous year (odds ratio (OR)=3.31, P<0.001) and was slightly higher among purchased cattle (37.6 per cent), than among cattle raised on the farm (29.1 per cent) (P=0.078). Seropositive cows were more likely than seronegative cows to have had a seropositive dam (OR=2.3, P=0.011), suggesting that congenital transmission contributed to about 56 per cent of the infections. Herds with a true seroprevalence above 10 per cent had more dogs on the farm, than herds with a lower prevalence (P=0.032). The ORs relating abortion to seropositivity in individual herds ranged from 0.7 to 19, indicating that some herds experienced few abortions caused by N caninum, while others experienced more abortions due to the organism. Overall, 38.7 per cent of the abortions were estimated to have been attributable to N caninum.

Efficacy of several serological tests and antigens for diagnosis of bovine brucellosis in the presence of false-positive serological results due to Yersinia enterocolitica O:9

ABSTRACT Yersinia enterocolitica O:9 bears a smooth lipopolysaccharide (S-LPS) of Brucella sp. O-chain A + C/Y epitopic structure and is a cause of false-positive serological reactions (FPSR) in standard tests for cattle brucellosis. Brucella S-LPS, cross-reacting S-LPSs representing several O-chain epitope combinations, Brucella core lipid A epitopes (rough LPS), Brucella abortus S-LPS-derived polysaccharide, native hapten polysaccharide, rough LPS group 3 outer membrane protein complexes, recombinant BP26, and cytosolic proteins were tested in enzyme-linked immunosorbent assays (ELISA) and precipitation tests to detect cattle brucellosis (sensitivity) and to differentiate it from FPSR (specificity). No single serological test and antigen combination showed 100% sensitivity and specificity simultaneously. Immunoprecipitation tests with native hapten polysaccharide, counterimmunoelectrophoresis with cytosolic proteins, and a chaotropic ELISA with Brucella S-LPS were 100% specific but less sensitive than the Rose Bengal test, complement fixation, and indirect ELISA with Brucella S-LPSs and native hapten or S-LPS-derived polysaccharides. A competitive ELISA with Brucella S-LPS and M84 C/Y-specific monoclonal antibody was not 100% specific and was less sensitive than other tests. ELISA with Brucella suis bv. 2 S-LPS (deficient in C epitopes), Escherichia hermannii S-LPSs [lacking the contiguous α-(1-2)-linked perosamine residues characteristic of Y. enterocolitica S-LPS], BP26 recombinant protein, and Brucella cytosolic fractions did not provide adequate sensitivity/specificity ratios. Although no serological test and antigen combination fully resolved the diagnosis of bovine brucellosis in the presence of FPSR, some are simple and practical alternatives to the brucellin skin test currently recommended for differential diagnosis.

Epidemiological pattern and risk factors associated with bovine viral-diarrhoea virus (BVDV) infection in a non-vaccinated dairy-cattle population from the Asturias region of Spain.

A survey of bovine viral-diarrhoea virus (BVDV) infection was carried out in a non-vaccinated cattle population from the Asturias region of Spain in 1997 to assess seroprevalence and identify risk factors associated with infection. Twenty-eight herds were included; 529 cows were bled. Information regarding the herd and each animal sampled were recorded through a personal interview with the farmer. The true prevalence was estimated to be 21%. According to the antibody-age profiles and the herd-management characteristics, no persistently infected animals were suspected at that time within the herds sampled. Random-effects logistic regression found two major factors associated with seropositivity: age and cow origin. Results suggested that BVDV infection could be controlled in that area by livestock-trade control (without vaccines). In addition, an increasing risk of abortion was not observed when cows were seropositive to both BVDV and Neospora caninum infections.

Salmonellosis in finishing pigs in spain: Prevalence, antimicrobial agent susceptibilities, and risk factor analysis

A herd-based survey of Salmonella in pigs was carried in a major pig producing region of Spain. Mesenteric lymph nodes were collected from the carcasses of 25 pigs from each of 80 herds at time of slaughter. Salmonella spp. were isolated from 31% of animals and 94% of herds. Within-herd prevalence ranged from 4 to 88%, with the prevalence in most herds being greater than 10%. A large diversity of Salmonella serotypes was found, with Typhimurium, 4,[5],12:i:-, and Rissen being the most prevalent. Two or more serotypes coexisted in 73% of the herds. Salmonella Typhimurium was present in 68% of the herds. Most (82%) of the Salmonella isolates belonged to serogroups targeted by enzyme-linked immunosorbent assay tests for pig salmonellosis. Resistance to at least one antimicrobial agent was detected in 73% of the strains, and one or more resistant strains were recovered from pigs in 93% of the herds. Antimicrobial agent resistance (AR) was more frequent among the most prevalent than it was among the rarer serotypes. Twenty-five multi-AR patterns were found. Resistance to three or more families of antimicrobial agents was found in 75% of AR strains. The finding that many of the herds yielded isolates of several multi-AR patterns indicates that Salmonella infections were acquired from multiple sources. High prevalence of Salmonella in herds was associated with lack of rodent control programs, herds from farms with only finishing pigs, herds managed by more than one full-time worker, herds for which the source of drinking water was not a city supply, and relatively long fattening times.

Epidemiology of subclinical salmonellosis in wild birds from an area of high prevalence of pig salmonellosis: phenotypic and genetic profiles of Salmonella isolates.

The epidemiology of subclinical salmonellosis in wild birds in a region of high Salmonella prevalence in pigs was studied. Three hundred and seventy‐nine faecal samples from 921 birds trapped in 31 locations nearby pig premises, and 431 samples from 581 birds of 10 natural settings far from pig farms were analysed for the presence of Salmonella spp. Positive samples were serotyped and analysed for antimicrobial resistance (AR). Phage typing and pulsed‐field gel electrophoresis (PFGE) on Salmonella Typhimurium isolates were also carried out. The overall proportion of Salmonella‐positive samples was 1.85% (95% CI = 0.93, 2.77). Salmonella isolation was positively associated with samples collected from birds in the proximity of a pig operation (OR = 16.5; 95% CI = 5.17, 52.65), and from non‐migratory (or short‐distance migration) birds (OR = 7.6; 95% CI = 1.20, 48.04) and negatively related to mostly granivorous birds (OR = 0.4; 95% CI = 0.15, 1.13). Salmonella Typhimurium was the most prevalent serotype and four different XbaI PFGE patterns were observed that matched the four phage types identified (U310, U311, DT164 and DT56). Only 20% of the strains showed multi‐AR. In three farms, a high degree of homogeneity among isolates from different birds was observed. These findings suggested that pig farms may act as amplifiers of this infection among wild birds, and the degree of bird density may have much to do on this transmission. Some of the Salmonella serotypes isolated from bird faeces were of potential zoonotic transmission and associated with AR. Monitoring salmonellosis in wild bird is advised.

Factors associated with seroprevalence to Mycobacterium paratuberculosis in small-ruminant farms in the Madrid region (Spain)

A cross-sectional study was conducted in a population of small ruminants in the Madrid region (Spain) to determine the Mycobacterium paratuberculosis seroprevalence and to identify farm factors possibly associated with paratuberculosis (PTB). Farming-management information and sera were collected from 60 sheep or goat flocks. The relationship between seropositivity and the variables in the questionnaire was assessed by unconditional logistic regression, followed by random-effects logistic regression analysis to adjust for overdispersion between herds. The seroprevalence to M. paratuberculosis was 11.7% (64 out of 546) using agar-gel immunodiffusion assay (AGID). According to the sensitivity and specificity of the AGID test the true prevalence could be as high as 44%. A herd size of between 200-400 head and the presence of foreign breeds and their crosses were significantly associated with seropositivity (OR = 4.05 and OR = 4.32, respectively). A higher replacement rate was also associated with seroprevalence to M. paratuberculosis (24.2% in positive herds vs. 18.1% in negative). All these three factors were related to more intensive management in the surveyed area. In contrast, membership of a professional livestock association appeared to be a protective factor against PTB (OR = 0.28). No variables related to veterinary assistance were associated with seroprevalence, probably reflecting the current lack of interest in PTB on the part of the animal-health administration and veterinary services in Spain.

Epidemiologic study of chlamydial infection in sheep farms in Madrid, Spain

Abstract A cross-sectional study was conducted in a population of sheep in Madrid (Spain) to assess seroprevalence and to identify risk factors for chlamydial infection. Information from 57 flocks was collected and 512 animals were sampled. The effects on the seroprevalence of several variables, such as farm management practices, farmer characteristics, animal health and veterinary services were evaluated using a random-effects logistic regression model. The seroprevalence to Chlamydia spp. was 50.5% (±4.5(95% CI)) using the Complement Fixation Test with a 1:32 cut-off titer. Fifty-five herds (96.5%) were positive. Three variables were associated with seroprevalence in the final model. Being a dairy animal was a risk factor (OR=2.40, P =0.067). Distance among farms greater than 500 m acted as protective factor (OR=0.55, * P =0.026). The third variable confirmed the endemic situation of chlamydiosis in the area: seropositive animals belonged to herds in which the observed percentage of abortions was greater than 5% (OR=2.40, * P =0.010).

Whole-cell and acellular pertussis vaccination programs and rates of pertussis among infants and young children

Background: The transition from a whole-cell to a 5-component acellular pertussis vaccine provided a unique opportunity to compare the effect that each type of vaccine had on the incidence of pertussis, under routine conditions, among children less than 10 years of age. Methods: Analyses were based on passive surveillance data collected between 1995 and 2005. The incidence of pertussis by year and birth cohort was compiled according to age during the surveillance period. We determined the association between vaccine type (whole-cell, acellular or a combination of both) and the incidence of pertussis using Poisson regression analysis after controlling for age (< 1 year, 1–4 years and 5–9 years) and vaccination history (i.e., partial or complete). Results: During 7 of the 11 years surveyed, infants (< 1 year of age) had the highest incidence of pertussis. Among children born after 1997, when acellular vaccines were introduced, the rates of pertussis were highest among infants and preschool children (1–4 years of age). Poisson regression analysis revealed that, in the group given either the whole-cell vaccine or a combination of both vaccines, the incidence of pertussis was lower among infants and preschool children than among school-aged children (5–9 years). The reverse was true in the group given only an acellular vaccine, with a higher incidence among infants and preschool children than among school-aged children. Interpretation: These results suggest that current immunization practices may not be adequate in protecting infants and children less than 5 years of age against pertussis. Altering available acellular formulations or adopting immunization practices used in some European countries may increase the clinical effectiveness of routine pertussis vaccination programs among infants and preschool children.

Sensitivity of the ISO 6579:2002/Amd 1:2007 Standard Method for Detection of Salmonella spp. on Mesenteric Lymph Nodes from Slaughter Pigs

ABSTRACT The ISO 6579:2002/Amd 1:2007 (ISO) standard has been the bacteriological standard method used in the European Union for the detection of Salmonella spp. in pig mesenteric lymph nodes (MLN), but there are no published estimates of the diagnostic sensitivity (Se) of the method in this matrix. Here, the Se of the ISO (SeISO) was estimated on 675 samples selected from two populations with different Salmonella prevalences (14 farms with a ≥20% prevalence and 13 farms with a <20% prevalence) and through the use of latent-class models in concert with Bayesian inference, assuming 100% ISO specificity, and an invA-based PCR as the second diagnostic method. The SeISO was estimated to be close to 87%, while the sensitivity of the PCR reached up to 83.6% and its specificity was 97.4%. Interestingly, the bacteriological reanalysis of 33 potential false-negative (PCR-positive) samples allowed isolation of 19 (57.5%) new Salmonella strains, improving the overall diagnostic accuracy of the bacteriology. Considering the usual limitations of bacteriology regarding Se, these results support the adequacy of the ISO for the detection of Salmonella spp. from MLN and also that of the PCR-based method as an alternative or complementary (screening) test for the diagnosis of pig salmonellosis, particularly considering the cost and time benefits of the molecular procedure.

Estimation of the diagnostic accuracy of the invA-gene-based PCR technique and a bacteriological culture for the detection of Salmonella spp. in caecal content from slaughtered pigs using Bayesian analysis.

The goal of this study was to estimate the accuracy of the invA‐gene‐based polymerase chain reaction (PCR) and a culture technique based on pre‐enrichment with buffered peptone water, three selective enrichment media (selenite, tetrathionate and Rappaport‐Vassiliadis broths) and four selective, solid media (Xylose‐Lysine‐Tergitol‐4, Salmonella/Shigella, Hekton‐Enteric and MacConkey), for the detection of Salmonella organisms from caecal samples from slaughter pigs. For this purpose a latent‐class (Bayesian) approach was used. Two hundred and three slaughtered pigs were used after grouping them into two groups of 96 and 107 animals. Sensitivity (Se) was estimated to be 56% (95% probability interval 40, 76) for culture and 91% (81, 97) for PCR. The specificity (Sp) of the PCR was 88% (80, 95) while the Sp of the culture had been considered 100% in the statistical analysis as all culture‐positive samples were confirmed by serotyping. PCR Se was not affected by the Salmonella serotypes present in the samples analysed. Accordingly, a minimum of 25.5% of the pigs was estimated to harbour Salmonella organisms in their faeces. It was concluded that bacteriology on caecal samples alone was a poor diagnostic method, and that the PCR method could be considered a cost‐effective alternative to culture in Salmonella monitoring programmes. However, given the moderate Sp of this molecular technique, PCR‐positive samples should be further confirmed through bacteriology.