Evaluation of ELISA coupled with Western blot as a surveillance tool for Trichinella infection in wild boar (original) (raw)
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Veterinary Parasitology, 2014
Please cite this article as: Cuttell, L., Gómez-Morales, M.A., Cookson, B., Adams, P.J., Reid, S.A., Vanderlinde, P.B., Jackson, L.A., Gray, C., Traub, R.J.,Evaluation of ELISA coupled with Western blot as a surveillance tool for Trichinella infection in wild boar (Sus scrofa), Veterinary Parasitology (2013), http://dx.
Indirect versus direct detection methods of Trichinella spp. infection in wild boar (Sus scrofa)
Parasites & Vectors, 2014
Background: Trichinella spp. infections in wild boar (Sus scrofa), one of the main sources of human trichinellosis, continue to represent a public health problem. The detection of Trichinella spp. larvae in muscles of wild boar by digestion can prevent the occurrence of clinical trichinellosis in humans. However, the analytical sensitivity of digestion in the detection process is dependent on the quantity of tested muscle. Consequently, large quantities of muscle have to be digested to warrant surveillance programs, or more sensitive tests need to be employed. The use of indirect detection methods, such as the ELISA to detect Trichinella spp. infections in wild boar has limitations due to its low specificity. The aim of the study was to implement serological detection of anti-Trichinella spp. antibodies in meat juices from hunted wild boar for the surveillance of Trichinella spp. infections.
Veterinary Parasitology, 2005
For the surveillance of trichinellosis, the digestion method is reliable but also labour intensive. The serological methods for the detection of Trichinella-specific antibodies using ELISA offer a sensitive and relatively specific alternative. For serological studies, sera or plasma from blood samples are the most common source of antibodies, but although the concentration of antibodies is approximately 10-fold lower, muscle fluid can be a good alternative particularly for testing of wildlife samples. In the present study, an indirect ELISA technique was evaluated on both sera and muscle fluids from experimentally infected foxes, pigs, and wild boars using both excretory/secretory (E/S) antigens and a synthetic glycan antigen, b-tyvelose. Although the synthetic antigen appears to be less sensitive than the E/S antigens, Trichinella-specific IgG antibodies were detected in both serum samples and muscle fluid samples from pigs, wild boars and foxes infected at levels which would be important for food safety or represent a significant reservoir for further transmission. #
Trichinella nematodes are the causative agent of trichinellosis, a meat-borne zoonosis acquired by consuming undercooked, infected meat. Although most human infections are sourced from the domestic environment, the majority of Trichinella parasites circulate in the natural environment in carnivorous and scavenging wildlife. Surveillance using reliable and accurate diagnostic tools to detect Trichinella parasites in wildlife hosts is necessary to evaluate the prevalence and risk of transmission from wildlife to humans. Real-time PCR assays have previously been developed for the detection of European Trichinella species in commercial pork and wild fox muscle samples. We have expanded on the use of real-time PCR in Trichinella detection by developing an improved extraction method and SYBR green assay that detects all known Trichinella species in muscle samples from a greater variety of wildlife. We simulated low-level Trichinella infections in wild pig, fox, saltwater crocodile, wild cat and a native Australian marsupial using Trichinella pseudospiralis or Trichinella papuae ethanol-fixed larvae. Trichinella-specific primers targeted a conserved region of the small subunit of the ribosomal RNA and were tested for specificity against host and other parasite genomic DNAs. The analytical sensitivity of the assay was at least 100 fg using pure genomic T. pseudospiralis DNA serially diluted in water. The diagnostic sensitivity of the assay was evaluated by spiking 10 g of each host muscle with T. pseudospiralis or T. papuae larvae at representative infections of 1.0, 0.5 and 0.1 larvae per gram, and shown to detect larvae at the lowest infection rate. A field sample evaluation on naturally infected muscle samples of wild pigs and Tasmanian devils showed complete agreement with the EU reference artificial digestion method (k-value = 1.00). Positive amplification of mouse tissue experimentally infected with T. spiralis indicated the assay could also be used on encapsulated species in situ. This real-time PCR assay offers an alternative highly specific and sensitive diagnostic method for use in Trichinella wildlife surveillance and could be adapted to wildlife hosts of any region.
Parasitology Research, 2008
The enzyme-linked immunosorbent assay (ELISA) method is recommended for farm surveillance programs and may be useful for epidemiological studies in wildlife or for establishing Trichinella-free areas. In this study, our interest was to compare the specificity and the time of seroconversion of excretory-secretory (E/S) antigens prepared from Trichinella spiralis. A group of eight pigs was inoculated with 500 T. spiralis larvae per animal, and blood sampling was performed at 3 and 4-day intervals during all experiments. The numbers of muscle larvae were determined in four different muscles groups. The larvae per gram burden shows that the most heavily parasitized muscles were the diaphragm [mean=43.7 larvae per gram (lpg)] and the tongue (mean=16.9 lpg). Antibody responses were detected by any of eight infected pigs of T. spiralis.
Prevalence of Trichinella spp. Infections in Hunted Wild Boars in Northern Iran
Iranian journal of public health, 2017
Trichinellosis is an important and neglected foodborne zoonotic infectious disease in worldwide. The most human outbreaks in recent years have been related to consumption of wild boar meat. This cross-sectional study determined the prevalence of Trichinella spp. infections in hunted wild boars in northern Iran. Thirty-five hunted wild boars were subjected in this study in 2015. All samples were examined by conventional artificial digestion method to detect of muscle larvae. Genomic DNA was extracted by phenol-chloroform method from isolated larvae. To identify the Trichinella species, a PCR-based method was applied using the internal transcribed spacer 2 (ITS2) and mitochondrial small-subunit ribosomal RNA (rRNA) gene sequences. The overall prevalence of Trichinella spp. infection was 5.7% (2/35, 95%CI= 0-13.4). The mean larval burdens in two positive samples were 0.05 and 6 larvae per gr tissue muscle, respectively. The PCR reaction, using specific primers, yielded two 367 bp and 1...
Seroprevalance of Trichinella spp. in wild boars (Sus scrofa) from Bihor county, western Romania
Helminthologia
SummaryThe wild boar (Sus scrofa) has a wide geographical distribution and can be an important source of Trichinella spp. infection in humans in Romania.The objective of this study was to identify the presence of Trichinella spp. in the wild boar population in Bihor County, Romania.Eighty four plasma and diaphragm samples, collected from wild boars, were included in this study. Artificial digestion, ELISA and Western blot were performed on these specimens. All diaphragm samples were negative for Trichinella larvae in artificial digestion, while in ELISA, 54 (64.2 %) plasma samples were positive and 6 (7.1 %) plasma samples were doubtful. Western blot was performed on 26 plasma samples from which only 6 (23.0 %) gave a positive result.Serological evidences indicate the presence of Trichinella spp. in wild boars from western Romania. Therefore, human consumers might be at risk to ingest Trichinella larvae, even in low numbers.
Journal of Helminthology, 2009
In Spain, trichinellosis represents a public health problem, with an average of five outbreaks per year, wild boar meat being the main source of infection. A trichinellosis survey (2007–2008 hunting campaign) was carried out on wild boars in the Toledo Mountains (south-western Spain, EU) in the context of a surveillance programme on wildlife diseases. A total of 2216 wild boars from different locations of the region were examined. The examination was carried out by veterinarians in the local abattoir (Matadero Municipal de Toledo). The positive samples were sent to the Department of Parasitology (Facultad de Farmacia, UCM) for experimental isolation and specific identification by inter-simple sequence repeat-polymerase chain reaction (ISSR-PCR). Using this technique we identified 17 isolates as Trichinella spiralis with an electrophoretic profile indistinguishable from the T. spiralis reference strain (ISS48). We confirmed that ISSR-PCR is a robust technique for the molecular identi...
Trichinella species circulating in wild boar (Sus scrofa) populations in Poland
International Journal for Parasitology: Parasites and Wildlife, 2013
Hunting in Poland has a long tradition and became more popular after 1990. Each year over 60,000 wild boar are hunted. Some of them may act as Trichinella carriers thus all carcasses of wild boar are systematically sampled in game-handling establishments as part of the post-mortem examination. The aim of the study was to determine the species of Trichinella and to evaluate the year to year differences in the occurrence of those species in the populations of wild boar in Poland. Samples for the study were provided by the Veterinary Inspection Service. Wild boar carcasses were examined using a digestion method. Only samples recognized as positive for Trichinella in these examinations were sent to the National Reference Laboratory (NRL) for confirmation of genus identity. Samples from 450 animals were obtained for the study (380 muscle samples and 70 larval isolates preserved in 90% ethyl alcohol). Tissue samples were digested to isolate larvae. Extracted larval DNA was amplified using a modified multiplex PCR protocol to identify the species of Trichinella. Five larvae from each sample were examined by PCR. The study revealed that Trichinella spiralis and Trichinella britovi are present in wild boar in Poland in a ratio of 3:1. Mixed infections with T. spiralis and T. britovi were found in 1% of the animals.