Comparative Evaluation of 2 In-Clinic Assays for Vector-Borne Disease Testing in Dogs

Vector-borne agents comprise medically important infections affecting dogs throughout much of the world. Sensitive detection of antibodies directed at tick-borne disease-causing organisms in dogs is diagnostically important for veterinarians, pets and their owners, and epidemiologically important for public health surveillance. The SNAP 4Dx Plus Test (IDEXX Laboratories, Inc., Westbrook, ME) identifies antibodies to or infection with multiple tick-borne pathogens and canine heartworm antigen in a single assay. Recently, VetScan FLEX4 Rapid Test (Abaxis, Inc., Union City, CA) was launched as a new assay to detect tick-borne pathogen antibodies and heartworm antigen. In the present study, we evaluated the comparative performance of SNAP 4Dx Plus (SNAP) and FLEX4 Rapid Test (FLEX4) using samples selected based on geographic distributions for canine vector borne diseases, including Borrelia burgdorferi (n = 105), Anaplasma phagocytophilum (160), Anaplasma platys (115), Ehrlichia canis (154), Ehrlichia ewingii (163), Ehrlichia chaffeensis (151) and Dirofilaria immitis (105). Canine vector borne diseases infection status was established for each sample by a combination of reference methods that included necropsy (D. immitis, heartworm disease), Western immunoblotting (B. burgdorferi), immunofluorescence assays (A. phagocytophilum and E. canis) and species-specific ELISAs (A. platys, E. canis, E. ewingii and E. chaffeensis). For comparisons among the 2 assays, samples were evaluated per the manufacturers’ instructions for each test kit.By testing each same sample set compared to the defined reference results, sensitivities differed substantially between SNAP and FLEX4, at 95.5 vs. 40.9%, respectively for B. burgdorferi, 97.1% vs. 61.4% for E. canis, 98.2% vs. 59.3% for E. ewingii, 64.3% vs. 35.7% for E. chaffeensis, 84.5% vs. 12.7% for A. phagocytophilum, 83.3% vs. 33.3% for A. platys, and 94.1% vs. 88.2% for D. immitis. Specificities for both rapid assay tests ranged from 98% to 100%.Based upon the comparative results derived from this study, the SNAP test was more sensitive than the FLEX4 test for detection of antibodies to all tick-borne pathogens and heartworm disease (Dirofilaria immitis) antigen in dogs.     

An investigation of the effects of intratesticular alfaxalone and lidocaine during castration in piglets

Objective

To determine whether intratesticular injection of an alfaxalone–lidocaine combination can induce anesthesia and provide a rapid recovery in piglets undergoing surgical castration.

Recreational sandboxes for children and dogs can be a source of epidemic ribotypes of Clostridium difficile

Different studies have suggested that the sand of public playgrounds could have a role in the transmission of infections, particularly in children. Furthermore, free access of pets and other animals to the playgrounds might increase such a risk. We studied the presence of Clostridium difficile in 20 pairs of sandboxes for children and dogs located in different playgrounds within the Madrid region (Spain). Clostridium difficile isolation was performed by enrichment and selective culture procedures. The genetic (ribotype and amplified fragment length polymorphism [AFLP]) diversity and antibiotic susceptibility of isolates was also studied. Overall, 52.5% (21/40) of samples were positive for the presence of C. difficile. Eight of the 20 available isolates belonged to the toxigenic ribotypes 014 (n = 5) and 106 (n = 2), both regarded as epidemic, and CD047 (n = 1). The other 12 isolates were non‐toxigenic, and belonged to ribotypes 009 (n = 5), 039 (n = 4), and 067, 151 and CD048 (one isolate each). Nevertheless, all isolates (even those of a same ribotype) were classified into different AFLP genotypes indicating non‐relatedness. In conclusion, our results revealed the presence of epidemic ribotypes of C. difficile in children's and dog's sandboxes located nearby, which constitutes a major health risk.     

Persistence of viral RNA in rabbits which overcome an experimental RHDV infection detected by a highly sensitive multiplex real-time RT-PCR

An internally controlled multiplex real-time RT-PCR using TaqMan probes and external standards for absolute RNA quantification was developed as a new diagnostic tool for the detection of rabbit haemorrhagic disease virus (RHDV). The test revealed a specificity of 100%, an analytical sensitivity of 10 copies/well and a linearity over a range from 10(1) to 10(10) copies. The viral loads in organs, leukocytes, sera and excretions of seropositive, convalescent rabbits which were overcoming an experimental infection with RHDV were determined using the validated assay. As a result, viral RNA was demonstrated and quantified for at least 15 weeks. Thus, a persistence of viral RNA after experimental infection of rabbits could be shown for the first time. In contrast, neither antigen nor infectious virus could be detected by antigen-ELISA, immunohistochemistry or experimental transmission. Therefore, further experiments are necessary to prove that the persistence of RNA is linked with the persistence of infectious virus particles.     

Evaluation of real-time RT-PCR assay for the routine intra vitam diagnosis of classical swine fever

The aim of the study was to evaluate the diagnostic quality of the real-time RT-PCR assay described by Hoffmann et al. [Hoffmann, B., Beer, M., Schelp, C., Schirrmeier, H., Depner, K., 2005. Validation of a real-time RT-PCR assay for sensitive and specific detection of classical swine fever. J. Virol. Methods 130, 36-44] for the routine intra vitam diagnosis of classical swine fever (CSF). We compared the assay with conventional diagnostic methods by using defined diagnostic material from an animal experiment with pigs showing different clinical forms of CSF. Compared to virus isolation and antigen ELISA an enhanced sensitivity of the real-time RT-PCR could be shown. We were able to detect all infected pigs regardless of the clinical course of CSF. CSF infection was detected already during the incubation period, during the entire clinical phase as well as at the beginning of convalescence when the first antibodies were detected and no virus could be isolated any more. In most cases, positive PCR results were obtained 2 days earlier than with virus isolation and 2-4 days earlier than with the antigen ELISA.     

Aspects on Hormonal Control of Normal and Induced Parturition in the Dog

Average length of gestation on the dog is 63 ± 2 days but may vary between 57-71 days due to the long period of receptivity at oestrus and the extended period of sperm survival in the female genital tract. In contrast to other domestic animals progesterone- and oestrogen concentrations are almost identical in pregnant and non pregnant bitches, except for their rapid decline immediately prior to parturition. Control of luteolysis still poorly understood. Experiments with indomethacin leading to a blockade of the prepartal PgF_2α increase, which commences with the decrease of progesterone, point toward a role of PgF_2α at this stage of pregnancy, which was extended for several days. At physiological conditions first visible signs of parturition were observed at peak-PgF_2α levels, 33.6 ± 17.6 hours after onset of luteolysis, which lasted over 16.8 ± 3.4 hours. Pulse-type releases of oxytocin were only observed after this point of time. To test for the effect of progesterone-withdrawal, four 51-57 days pregnant bitches were treated with the antiprogestin RU 38486 which inhibits the activity of progesterone at the receptor level. In all dogs first visible signs of parturition were observed 33.5 ± 7.5 hours after onset of treatment. However, the process of parturition came to an end after cervical opening and totally only one puppy was born. Different to a normal parturition no increase of PgF_2α was observed. Relaxin levels were not influenced by treatment. These observations suggested that treatment with antiprogestin followed by PgF_2α might be an adequate method to induce parturition in the dog; first experiences seem to confirm this conclusion.     

Evaluation of extreme resistance genes of Potato virus X (Rx1 and Rx2) in different potato genotypes

Journal of Plant Diseases and Protection - Potato (Solanum tuberosum L.) is considered as one of the most important staple foods in the world. Classical breeding for resistance to pests, pathogens...     

Genotypic differences in storage losses of sugar beet – causes and indirect criteria for selection

To improve the storability of sugar beets, this study aimed at determining reasons for genotypic variability in sugar losses and invert sugar accumulation during storage, and at identifying indirect criteria to select for varieties with low storage losses prior to storage. In 2011 and 2012, 18 genotypes, and in 2012 and 2013, six genotypes cultivated at two locations were stored for 8 and 12 weeks at 8°C under controlled conditions. The same 18 genotypes were grown under stress conditions in Spain in 2012/2013. Sugar losses were closely correlated with the invert sugar accumulation after storage. Genotypic differences in storage losses were primarily caused by differences in the level of infestation with microorganisms. The invert sugar accumulation was lower for genotypes with high marc concentration before storage, pointing to a non‐specific resistance. Additionally, the sugar concentration in dry matter before storage, and the invert sugar concentration after cultivation under stress conditions correlated with the invert sugar concentration after storage. These parameters are therefore suggested as criteria to select for improved storability of sugar beet genotypes.