Comparison of the Q-fever complement fixation test and two commercial enzyme-linked immunosorbent assays for the detection of serum antibodies against Coxiella burnetii (Q-fever) in ruminants: Recommendations for use of serological tests on imported animals in New Zealand

Abstract

AIM: To make valid recommendations on the use of serological test methods for the detection of serum antibodies in ruminants against Coxiella burnetii (Q-fever), by comparing the performance of the complement fixation test (CFT) and two ELISA, and by identifying reasons for discrepancies between the test methods.

METHODS: A total of 73 serum samples from infected cattle, 69 from infected goats, and 100 samples from non-infected cattle and 57 samples from non-infected sheep, as well as 95 samples from infected cattle herds (mix of seropositive and seronegative samples), were tested using the CFT, the IDEXX ELISA (I-ELISA) and the Pourquier ELISA (P-ELISA). A mixed panel of 12 serum samples from sheep from inter-laboratory proficiency testing (proficiency panel) was also tested using the CFT and both ELISA, and further investigated using IgG- and IgM-specific ELISA.

RESULTS: Generally, the two commercial ELISA were more sensitive than the CFT for the detection of infected ruminants. Good agreement between ELISA for positive and negative results was found for samples from the infected herd, while results for the positive panels varied between the two ELISA. For the total of the positive serum panels, the I-ELISA detected 95% of samples as positive or suspicious, while the P-ELISA detected only 81%. In the P-ELISA, more samples were considered suspicious (18%) than in the I-ELISA (14%). All sera from noninfected sheep and cattle tested negative in the serological test methods employed, except for one positive sample from a sheep in the P-ELISA. Further investigation revealed that a CFT-positive but ELISA-negative result was due to high IgM and low IgG reactivity.

CONCLUSIONS: The two commercial ELISA were more sensitive than the CFT in all panels from infected ruminants. However, they could only detect IgG. The I-ELISA should be the serological test method of choice for cattle, sheep and goats for import testing of animals into New Zealand because it was more sensitive than the P-ELISA and was equally specific to the PELISA and the CFT. For other animal species, such as deer and camelids, the CFT should still be used since none of the ELISA has been evaluated for these species. This study has shown that the two commercial ELISA will detect the majority of infected ruminants but may miss animals that have not developed an IgG response.     

Diarrhea and loss of production on Dutch dairy farms caused by the Schmallenberg virus

At the end of August and the first two weeks of September 2011 dozens of veterinary practitioners reported to GD Veekijker (Animal Health Service) several dairy herds with cows with sudden decreased milk production, watery diarrhea and sometimes fever. In the beginning these reports came from the Eastern region of the Netherlands, after that also from the other three regions. The percentages of diseased herds per veterinary practice varied from a few till dozens per cent. Extensive bacteriological, virological and parasitological testing of the feces of sick cows did not reveal an infectious cause of the clinical problems. Recently, 50 stored blood samples of clinically diseased cattle were tested for the Schmallenbergvirus using a PCR, and 36% (18/50) tested positive. A large group of control cows (n=115) was also tested with the PCR and all cattle tested negative. Likely the Schmallenbergvirus was the primary cause of the clinical symptoms in the Dutch dairy herds. Further research will be done to confirm this.     

Airborne transmission of bovine herpesvirus 1 infections in calves under field conditions

A small scale transmission experiment was performed with bovine herpesvirus 1 (BHV1) in a cattle population under field conditions. 10 calves were housed under strict hygienic conditions, with a distance of 4m between each calf. Five calves were experimentally infected with BHV1, two calves with strain Harberink and three with strain Lam, respectively. Experimentally infected calves were placed at 4 m distance from five susceptible sentinel calves. Airborne transmission to sentinel calves was detected using virus isolation and BHV1 specific polymerase chain reactions in samples of nasal fluids, and BHV1 specific antibodies in serum samples. Strain Harberink was hardly transmitted to sentinel calves, whereas strain Lam was transmitted to all sentinels. Estimating the rate of transmission per day, the total number of calves infected by one (strain Lam) infected calf was 1.18. Comparing this estimated transmission ratio between cattle at a distance of 4 m to the estimated transmission ratio R of BHV1 in susceptible commingled cattle reported before, the effect of the factor distance on the transmission ratio could be calculated. Extrapolating these results, a distance of 4.4 m between cattle populations would be necessary to reduce transmission for this strain to R<1.     

The results of using faecal culture as confirmation test of paratuberculosis-seropositive dairy cattle

A total of 15,822 cattle aged 3 years and older, belonging to 378 randomly selected herds, were tested for paratuberculosis using an absorbed enzyme-linked immunosorbent assay (ELISA); 3.3% tested positive. This percentage was lowest for the group of cattle aged 3-4 years (2.3%) and highest for cattle with the age of 5-6 years (4.5%). The mean Sample to Positive (S/P) ratio of seropositive cattle vaccinated against paratuberculosis was higher (0.75 +/- 0.33) than that of seropositive, non-vaccinated cattle (0.58 +/- 0.26). Faecal samples of 422 ELISA-positive cattle were cultured for the presence of Mycobacterium avium subsp. paratuberculosis, 12% of these were contaminated. The percentage of non-contaminated samples with positive culture results was 17.3%, with a substantial difference between vaccinated (1.7%) and non-vaccinated cattle (20.2%). Of the positive cultures, the number of colonies varied from 1-10 (22% of cultures), 11-100 (22%), to more than 100 (55%). The percentage of ELISA-positive, non-vaccinated cattle tested culture-positive was positively correlated with the magnitude of the S/P ratio. This percentage varied from 12% (S/P ratio 0.3-0.5) to 58% (S/P ratio > 1.1), a result that might have implications for interpretation of the test. In this study, the percentage of ELISA-positive cattle with positive faecal culture results was limited and these individuals were mostly moderate to heavy shedders.     

Genetic analysis of Tunisian fig (Ficus carica L.) cultivars using amplified fragment length polymorphism (AFLP) markers

Genetic diversity of forty fig cultivars collected from five regions in Tunisia was investigated using amplified fragment length polymorphism (AFLP). A total of 342 reproducible bands amplified with six AFLP primer combinations were obtained. The high percentage of polymorphic bands (%PB) of 97.5 and the resolving power (Rp) collective rate value of 143 were scored. In addition, the polymorphism information content (PIC) values varied from 0.61 to 0.87 with an average of 0.77. Although cluster (UPGMA) and principal components analyses indicate that the cultivars’ clustering made independently both from the geographical origin, horticultural classifications and/or from the sex of trees. In addition, the observed variation suggests considerable differentiation among fig cultivars. The present data supports the common origin of the fig cultivars. Analysis of molecular variance (AMOVA) revealed that average Φ_ST value overall loci was 0.026, and the overall distribution pattern of molecular variation indicated that about 97.43% of the total variance was accounted by the within-region variance component. The remaining 2.5% (P < 0.001) of the variation was founded among cultivars of the prospected regions. Our results proved that AFLP markers are useful for germplasm discrimination as well as for investigation of fig patterns variation. The information may be useful to define conservation management program.     

Occurrence of Biscogniauxia mediterranea in cork oak stands in Tunisia

Phytoparasitica - Cork oak decline and mortality are increasingly prevalent in the Mediterranean basin. This require particular attention due to the great economic value of this plant species....     

Potassium uptake efficiency of two pear cultivars and leaf concentration at deficiency symptoms appears

Two pear cultivars (Meski Ahrech and Alexandrine) grown in pot culture under normal daylight conditions in a greenhouse were submitted to potassium deficiency treatments. The potassium-free treatment decreased leaf K concentration, relative water content, SPAD, and gas exchange comparatively to the control. “Alexandrine” appeared more sensitive to potassium deficiency due to the main decrease in stomatal conductance (gs), photosynthesis (A) and transpiration (E) compared to “Meski Ahrech” that reduced only carbon assimilation (A). After 42 days of treatments, leaf K concentration in “Alexandrine” was 0.75% DM compared to 1.15% DM in “Meski Ahrech”, which appeared to be more efficient in potassium uptake.