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Capripox viruses are the causative agents of important animal diseases in cattle (Lumpy Skin Disease), sheep (Sheeppox) and goats (Goatpox) with severe socio-economic impact in case of wide scale outbreaks. Therefore there is a constant need for adequate diagnostic tools. The assays must be fit-for-purpose to identify the virus quickly and correctly and to be useful for surveillance and monitoring at different stages of an epidemic. Different diagnostic performance characteristics are required depending on the situation and the test purpose. The need for high throughput, high specificity/sensitivity and the capability for differentiating field virus strains from vaccine strains drives the development of new and better assays preferably with an advantageous cost-benefit balance. This review aims to look at existing and new virological and serological diagnostic tools used in the control against diseases caused by Capripox viruses.

Within the framework of the national voluntary eradication program for Bovine alphaherpesvirus 1 (BoHV1) in France, the proportion of certified-free herds which experienced no more than two positive animals (termed singleton reactors) steadily increased to reach up to 95% in 2015. The aim of this study was to collate and evaluate serological data to gain insight into these epidemiological questionable BoHV1 seropositive animals. Preliminary evaluation of the performances of BoHV1 ELISA kits using a collection of 997 field sera with well-defined status revealed a relatively low specificity of the two gB blocking ELISAs most used in France for confirmatory testing (93.2% and 97.5% for gB-IDVet and gB-Idexx, respectively). In both ELISAs, the suboptimal specificity was associated with the presence of antibodies against BoHV2. Reassessment of the cut-offs led to a specificity and a sensitivity higher than 99.3%. Consequently, a comprehensive analysis of gB-positive sera from 2551 singleton reactors was performed by using gB ELISAs with optimized cut-offs, combined with viral neutralization test (campaign 2014-2015) or gE ELISA (campaign 2015-2016). Fifty percent of the 728 sera collected in 2014-2015 reacted below the optimized cut-offs in both gB ELISAs. Analysis of new blood samples collected at a minimum 6-week interval showed that these weak-positive reactions did not increase with time and could not be confirmed by confirmatory tests. Among the 1823 sera collected in 2015-2016, only 84 samples tested positive by gE ELISA, most of them corresponding to sera with reactivity above the optimized cut-offs in gB ELISAs. Screening for BoHV2 antibodies revealed a significantly increased prevalence among herds with singleton reactors, compared with the between-herd prevalence in French cattle herds. Altogether, these results provided suitable analytical strategies to limit the occurrence of false-positive BoHV1 reactions and inappropriate withdrawal of the BoHV1-free status, without alteration of diagnostic costs and reliability of eradication programs.

Low sensitivity (Se) of diagnostic tools is often mentioned as a major problem in the control of paratuberculosis (PTB) and much effort is put into the improvement of these tests. The hypothetical perspectives of improving the Se of a milk-antibody ELISA (hereafter: milk-ELISA) used in test-&-cull strategies against PTB in dairy cattle were investigated by simulations. The current Se varies between 10 and 80%, increasing with increasing lactation stage, parity and infection stage. We simulated the effects on a dairy herd's production of improving this Se to 80% (independent of these factors) and assumed no concomitant decrease in specificity. By using a PTB model called PTB-Simherd, 12 scenarios were simulated to study three test-&-cull strategies in each of four herds with 200 dairy cows. To show the maximal effect of using test-&-cull with such an improved test we simulated three strategies: (1) no testing, (2) testing with milk-ELISA test with the current Se and culling of positive cows immediately and (3) testing with milk-ELISA test with a Se improved to 80% and culling positive cows immediately. The four herds were defined by a moderate (25%) or high (80%) initial true within-herd prevalence (including young stock), and a poor or good heat-detection success of 40 or 60%. We assumed that these factors influenced the effects of improving the Se of the milk-ELISA. Management both concerning calf management and in general was specified to represent a typical Danish herd. Using an improved milk-ELISA was predicted to reduce the prevalence of PTB more effectively than the current ELISA, and over 10 years bring the production of a herd with moderate initial prevalence up to a production level comparable to a non-infected herd (unlike if the current ELISA had been used). In a herd with high initial prevalence (80%) milk production was increased more by using the improved milk-ELISA, but after 10 years the replacement rate was still very high causing problems with having enough recruitment animals—especially in high-prevalence herds with poor reproductive performance. Economically important measurements in all four herds benefited from the improvement of the test over a 10-year period. However, in the first 3–5 years the improved test would be more expensive to use than the current test, due to increased replacement (reduced net annual revenue per cow €15 on average) but after that, net annual revenue increased continuously; after 10 years it was €70–90 higher, than if the current milk-ELISA was used. Also, the milk-ELISA test with its current Se seemed to be profitable already after 2 years in high-prevalence herds using a test-&-cull strategy based on the milk-ELISA alone.

Veterinary inspection in slaughterhouses allows for the detection of macroscopic lesions reminiscent of bovine tuberculosis, but the presence of Mycobacterium bovis must be confirmed by laboratory methods. This study aimed at comparing the performances of the standard diagnostic tools used to identify M. bovis in tissue specimens sampled from suspicious animals. During a two years period, 1390 cattle were inspected at the Machachi abattoir in the Mejia canton Ecuador. A total of 33 animals with granulomatous lesions were detected, representing 2.33% (16/687) and 2.42% (17/703) animals examined in 2007 and 2008, respectively. Ninety-four tissue specimens were sampled and screened for the presence of mycobacteria. Acid-fast bacilli were identified in one third of the suspicious cattle (11/33) and suggestive microscopic lesions in 27.3% (9/33) of the samples examined by direct microscopy and histopathology, respectively. Culturing on Stonebrink medium and 16S-rRNA-based polymerase chain reaction (PCR) yielded 36.4% (12/33) and 27.3% (9/33) of positives, respectively. Compared to culture, other diagnostic procedures displayed a lower sensitivity, with 56.5% for PCR, and 43.5% for direct microscopy and histopathology; however, the specificity was higher (94.4% for PCR and microscopy, and 97.2% for histopathology). We conclude that reliable post-mortem laboratory testing either requires the combination of a set of available diagnostic tools or necessitates the development of improved new-generation tools with better sensitivity and specificity characteristics.