[Show abstract][Hide abstract] ABSTRACT: Background
Parasites of the genus Trichinella are zoonotic nematodes common in carnivores throughout the world. We determined the prevalence and species of Trichinella infections in Florida panthers (Puma concolor coryi).Methods
Tongues from Florida panthers were collected at necropsy and examined by pepsin-HCl artificial digestion for infection with Trichinella spp. DNA was extracted from larvae and multiplex PCR using Trichinella species-specific primers was used to genotype the worms.Results Trichinella spp. larvae were detected in 24 of 112 (21.4%; 14.6%¿30.3%) panthers. Sixteen of the panthers (14.3%) were infected with T. pseudospiralis, 1 (0.9%) was infected with T. spiralis, and 2 (1.8%) had mixed infections of T. pseudospiralis and T. spiralis. Trichinella spp. larvae from 5 panthers were not identified at the species level due to degraded DNA.Conclusions
This is the highest prevalence of T. pseudospiralis detected in North America up to now and suggests the Florida panther is a key mammalian reservoir of this parasite in southern Florida. Trichinella pseudospiralis can infect both mammals and birds indicting the source of infection for Florida panthers could be broader than believed; however, birds represent a small percentage (0.01%) of the cat¿s diet. Since wild pigs (Sus scrofa) can be parasitized by both T. pseudospiralis and T. spiralis and that these swine can comprise a large portion (~40%) of a panther¿s diet in Florida, we believe that Florida panthers acquired these zoonotic parasites from feeding on wild pigs.
[Show abstract][Hide abstract] ABSTRACT: Serology to monitor Trichinella spp. infection in pigs reared in controlled system has been claimed as a possible diagnostic tool. However, no international biological standards or reference materials exist to validate in house tests or commercial kits, and to improve the inter-laboratory comparability for the serological detection of anti-Trichinella IgG in pigs. In this work, potential reference sera have been prepared from four experimentally infected pigs. Sera were tested, aliquot, lyophilized, and maintained at +4 °C. Since one of the prerequisites for the development of any reference material is to plan and execute stability studies, isochronous studies for short and long term stability testing were carried out to evaluate the possible degradation effects of transportation and storage. The stability of the lyophilized serum samples at +4 °C, was arbitrarily assumed. For the short term stability study, two units were stored at -20 °C, +4 °C, +20 °C, and +50 °C for 0, 1, 2 and 4 weeks, and then tested in duplicate. For the long term stability study, the same number of units and replicates per unit were stored at -80 °C, -20 °C, and +4 °C, for 0, 6, 12, 18 and 24 months. In both studies, unit samples were selected randomly and tested on the same day under repeatability conditions. The linear regression versus time for each serum at each studied temperature was analyzed and then slopes were tested for significance. Further, uncertainty of the short and long term stability was calculated for a shelf life period of one week and three years, respectively. For all sera but one, and for all the studied temperatures but +50 °C, the data from the short term stability study indicate the absence of a significant trend that would hint at degradation. The slopes of the regression lines did not significantly vary from zero. Even if the uncertainty of the short term stability was variable among serum samples, the rate of degradation was considered acceptable. For the long term stability, slopes of the regression lines of two serum samples significantly varied from zero, indicating a trend of possible degradation during storage. The percentage of degradation deducted from the uncertainty of the long term study varied; however, two serum samples showed the lower rate of degradation at all the assayed temperatures. The most suitable temperatures for dispatching serum samples are -20 °C, +4 °C and +20 °C; whereas, -20 °C and -80 °C are suitable temperatures for serum storage.
[Show abstract][Hide abstract] ABSTRACT: In the present study we sequenced or re-sequenced, assembled and annotated 15 mitochondrial (mt) genomes representing the 12 currently recognised taxa of Trichinella using a deep sequencing-coupled approach. We then defined and compared the gene order in individual mt genomes (∼ 14 to 17.7 kb), evaluated genetic differences among species/genotypes and re-assessed the relationships among these taxa using the mt nucleic acid or amino acid sequence data sets. In addition, a rich source of mt genetic markers was defined that could be used in future systematic, epidemiological and population genetic studies of Trichinella. The sequencing-bioinformatic approach employed herein should be applicable to a wide range of eukaryotic parasites.
International Journal for Parasitology 09/2014; · 3.40 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Abstract Trichinella spiralis and Trichinella britovi are the two most common species of the genus Trichinella persisting in the European wildlife. To investigate the spatial distribution of these Trichinella spp. and the factors influencing their circulation in Hungary, 3304 red foxes (Vulpes vulpes) and 0.29 million wild boars (Sus scrofa) were tested for Trichinella sp. infection in Hungary from 2006 to 2013. Trichinella spp. larvae from 68 (2.06%) foxes and 44 (0.015%) wild boars were identified by a multiplex PCR as T. britovi or T. spiralis. The locality of origin of foxes and wild boars were recorded in a geographic information system database. There was no correlation between environmental parameters in the home range of foxes and wild boars and the T. spiralis larval counts, but there was a positive correlation between the boundary zone of Hungary and T. spiralis infection (P < 0.0001; odds ratio: 24.1). These results indicate that the distribution of T. spiralis in the Hungarian wildlife is determined by the transborder transmission of the parasite from the surrounding endemic countries. Multiple regression analysis was performed with environmental parameter values and T. britovi larval counts. Based on the statistical analysis, non-agricultural areas (forests, scrubs, herbaceous vegetation and pastures) and the mean annual temperature (P < 0.0001; odds ratios: 9.53 and 0.61) were the major determinants of the spatial distribution of T. britovi in Hungary. The positive relationship with non-agricultural areas can be explained by the generalist feeding behaviour including scavenging of foxes in these areas. The negative relationship with the mean annual temperature can be attributed to the slower decomposition of wildlife carcasses favouring a longer survival of T. britovi larvae in the host carrion and to the increase of scavenging of foxes.
[Show abstract][Hide abstract] ABSTRACT: Trichinella spiralis and Trichinella pseudospiralis exhibit differences in the host-parasite relationship such as the inflammatory response in parasitized muscles. Several studies indicate that matrix metalloproteinases (MMPs) represent a marker of inflammation since they regulate inflammation and immunity. The aim of this study was to evaluate the serum levels of gelatinases (MMP-9 and MMP-2) in mice experimentally infected with T. spiralis or T. pseudospiralis, to elucidate the involvement of these molecules during the inflammatory response to these parasites. Gelatin zymography on SDS polyacrilamide gels was used to assess the serum levels and in situ zymography on muscle histological sections to show the gelatinase-positive cells.In T. spiralis infected mice, the total MMP-9 serum level increased 6 days post infection whereas, the total MMP-2 serum level increased onward. A similar trend was observed in T. pseudospiralis infected mice but the MMP-9 level was lower than that detected in T. spiralis infected mice. Significant differences were also observed in MMP-2 levels between the two experimental groups. The number of gelatinase positive cells was higher in T. spiralis than in T. pseudospiralis infected muscles. We conclude that MMP-9 and MMP-2 are markers of the inflammatory response for both T. spiralis and T. pseudospiralis infections.This article is protected by copyright. All rights reserved.
[Show abstract][Hide abstract] ABSTRACT: The wild boar is an important source of trichinellosis for people in European countries as a large number of hunted animals escape veterinary control. In November 2012, uncooked sausages made with meat from wild boar were consumed by 38 persons in a village of the Lucca province (Tuscany region, Italy). Of them, 34 were serologically positive, 32 developed clinical signs and symptoms of trichinellosis, and two were asymptomatic. Trichinella britovi larvae were detected in vacuum-packed sausages made with the same batch of sausages consumed raw which had been prepared with meat from wild boar hunted in the Lucca province. As no case of trichinellosis had been reported in this region during the last 20 years, the regional public health authority considered the risk for this zoonosis to be negligible and put in place a surveillance programme on Trichinella spp. in indicator animals (mainly foxes and including wild boar for private consumption), by testing only a percentage of heads. The experience from this outbreak shows that the definition of a region with a negligible risk for Trichinella infection is not applicable to wild boar and stresses the need to test all Trichinella-susceptible wild animals intended for human consumption and to implement risk communication to consumers and hunters.
Zoonoses and Public Health 08/2014; · 2.09 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The objective of this study was to assess the presence of anti-Trichinella IgG in the serum of persons from ethnic minorities from northwest Vietnam with clinical signs and symptoms that are compatible with trichinellosis. A total of 645 persons were enrolled, of which 200 people lived in two villages where outbreaks of human trichinellosis had been documented in 2004 and 2008, and 445 people who were hospitalized in the Dien Bien and Son La provincial hospitals without a definitive diagnosis. Presence of anti-Trichinella IgG was demonstrated in serum samples by a standardized Enzyme-linked Immunosorbant Assay (ELISA); positive serum samples were subjected to Western Blot (WB) for confirmation. Seven (3.5%; 95% CI: 1.4 - 7.1) persons from the villages and seven (1.6%; 95% CI: 0.6 - 3.2) hospitalized patients, tested positive by both ELISA and WB. Fever (N=13), eosinophilia (N=12), myalgia (N=9), facial oedema (N=9) and leukocytosis (N=8) were the most common clinical signs and symptoms in the serologically positive persons. The concomitant occurrence of facial oedema and myalgia among the enrolled persons from the villages, accounted for 75% of the positive predictive value (PPV) and 99.5% of the negative predictive value (NPV), suggesting that they could be used for suspecting trichinellosis when serology is not available. The high prevalence (1.6 - 3.5%) of anti-Trichinella IgG in persons from Vietnamese provinces where T. spiralis is circulating in pigs strongly supports the need to develop control programmes to eliminate the infection from pigs and for consumers' education and protection.
[Show abstract][Hide abstract] ABSTRACT: Several outbreaks of trichinellosis associated with the consumption of raw pork have occurred in Laos since 2004. This cross-sectional study was conducted in four provinces of northern Laos to investigate the seroepidemiology of trichinellosis in the human population and determine the prevalence and species of Trichinella infection in the domestic pig population. Serum samples and questionnaire data were obtained from 1419 individuals. Serum samples were tested for Trichinella antibodies by ELISA using larval excretory-secretory (ES) antigens and a subset of 68 positive samples were tested by western blot. The seroprevalence of Trichinella antibodies was 19.1% (95% confidence interval (CI) = 17.1-21.1%). The risk of having antibodies detected by ELISA using ES antigens increased with age, being of Lao-Tai ethnicity, living in Oudomxay province and being male. Tongue and diaphragm muscle samples were collected from 728 pigs and tested for Trichinella larvae by the artificial digestion method. Trichinella larvae were isolated from 15 pigs (2.1%) of which 13 were identified as T. spiralis by molecular typing; the species of the two remaining isolates could not be determined due to DNA degradation. Trichinella spp. are endemic in the domestic environment of northern Laos and targeted preventative health measures should be initiated to reduce the risk of further outbreaks occurring.
[Show abstract][Hide abstract] ABSTRACT: Trichinellosis is a cosmopolitan foodborne disease that may result in severe health disorders and even death. Despite international awareness of the public health risk associated with trichinellosis, current data on its public health impact are still lacking. Therefore we assessed, for the first known time, the global burden of trichinellosis using the Disability-Adjusted Life Year (DALY) metric. The global number of DALYs due to trichinellosis was estimated to be 76 per billion persons per year (95% credible interval (CrI): 38-129). The World Health Organization (WHO) European Region was the main contributor to this global burden, followed by the WHO region of the Americas and the WHO Western Pacific region. The global burden of trichinellosis is much lower than that of other foodborne parasitic diseases and is in sharp contrast to the high budget allocated to prevent the disease in many industrialized countries. To decrease the uncertainty around the current estimates, more knowledge is needed on the level of underreporting of clinical trichinellosis in different parts of the world.
International Journal for Parasitology 06/2014; · 3.40 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: 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.
Two tests were used, ELISA for the initial screening test, and a specific and sensitive Western blot (Wb) as a confirmatory test. The circulation of anti-Trichinella IgG was determined in hunted wild boar muscle juice samples in 9 provinces of 5 Italian regions.
From 1,462 muscle fluid samples, 315 (21.5%, 95% C.I. 19.51-23.73) were tested positive by ELISA. The 315 ELISA-positive muscle fluid samples were further tested by Wb and 32 (10.1%, 95% C.I. 7.29-13.99) of these were positive with a final seroprevalence of 2.2% (95% C.I 1.55-3.07; 32/1,462). Trichinella britovi larvae were detected by artificial digestion in muscle tissues of one (0.07%, 95%C.I. 0.01-0.39) out of the 1,462 hunted wild boars. No Trichinella spp. larvae were detected in Wb-negative wild boar. From 2006 to 2012, a prevalence of 0.017% was detected by muscle digestion in wild boar hunted in the whole Italian territory.
The combined use of both serological methods had a sensitivity 31.4 times higher than that of the digestion (32/1,462 versus 1/1,462), suggesting their potential use for the surveillance of the Trichinella spp. infection in wild boar populations.
[Show abstract][Hide abstract] ABSTRACT: The 14-3-3s are a family of dimeric evolutionary conserved pSer/pThr binding proteins that play a key role in multiple biological processes by interacting with a plethora of client proteins. Giardia duodenalis is a flagellated protozoan that affects millions of people worldwide causing an acute and chronic diarrheal disease. The single giardial 14-3-3 isoform (g14-3-3), unique in the 14-3-3 family, needs the constitutive phosphorylation of Thr214 and the polyglycylation of its C-terminus to be fully functional in vivo. Alteration of the phosphorylation and polyglycylation status affects the parasite differentiation into the cyst stage. To further investigate the role of these post-translational modifications, the crystal structure of the g14-3-3 was solved in the unmodified apo form. Oligomers of g14-3-3 were observed due to domain swapping events at the protein C-terminus. The formation of filaments was supported by TEM. Mutational analysis, in combination with native PAGE and chemical cross-linking, proved that polyglycylation prevents oligomerization. In silico phosphorylation and molecular dynamics simulations supported a structural role for the phosphorylation of Thr214 in promoting target binding. Our findings highlight unique structural features of g14-3-3 opening novel perspectives on the evolutionary history of this protein family and envisaging the possibility to develop anti-giardial drugs targeting g14-3-3.
PLoS ONE 03/2014; 9(3):e92902. · 3.53 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Each year, millions of pigs worldwide are tested for Trichinella spp. at slaughterhouses with negative results. Yet, thousands of people acquire trichinellosis by consuming pork. So, where is the problem? Testing for Trichinella spp. is often performed on the 'wrong' animals; while the parasites are mainly circulating in backyard and free-ranging pigs, herds kept under controlled management conditions are the ones tested. Veterinary services should: (i) introduce a risk-based surveillance system for Trichinella by documenting the control of housing conditions and feedstuff sources, and (ii) introduce a capillary network of field laboratories for monitoring the parasites in free-ranging and backyard pigs. Investment of funds into the education of farmers, hunters, and consumers should be a priority for public health services.
Trends in Parasitology 12/2013; · 6.22 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Trichinella infections in humans and pigs have been documented in Greece since 1945 and a high prevalence of infection in pigs occurred in the 1950s. Up to 1984 only sporadic infections in humans were documented, and this zoonosis was not considered as a public health problem until 2009 when a human outbreak caused by the consumption of pork from an organic pig farm occurred. In the present study, we describe the re-emergence of Trichinella spp. infections in free-ranging pigs from organic farms of 3 counties (Dramas, Evros and Kavala) in Northern-Eastern Greece during the period 2009-2012. Totally 37 out of 12,717 (0.29%) free-ranging pigs which were tested during the period in question, were positive for Trichinella spp. larvae. The etiological agent was identified as Trichinella britovi. The average larval burden was 13.7 in the masseter, 6.2 in the foreleg muscles and 7.5 in the diaphragm. The 37 positive animals originated from seven free range pig farms. The practice of organic pig production systems in Greece has grown in popularity over the last years due to the increasing interest of consumers for products considered as traditional. However, this type of pig production increases the risk for Trichinella spp. infections, since animals can acquire the infection by feeding on carcasses or the offal of hunted or dead wild animals. The awareness and education of hunters and farmers is extremely important to reduce the transmission among free ranging pigs and the risk for humans.
[Show abstract][Hide abstract] ABSTRACT: The 14-3-3s are small acidic cytosolic proteins that interact with multiple clients and participate in essential cellular functions in all eukaryotes. Available structural and functional information about 14-3-3s is largely derived from higher eukaryotes, which contain multiple members of this protein family suggesting functional specialization. The exceptional sequence conservation among 14-3-3 family members from diverse species suggests a common ancestor for 14-3-3s, proposed to have been similar to modern 14-3-3ε isoforms. Structural features of the sole family member from the protozoan Giardia duodenalis (g14-3-3), are consistent with this hypothesis, but whether g14-3-3 is functionally homologous to the epsilon isoforms is unknown. We use inter-kingdom reciprocal functional complementation and biochemical methods to determine whether g14-3-3 is structurally and functionally homologous with members of the two 14-3-3 conservation groups of the metazoan Drosophila melanogaster. Our results indicate that although g14-3-3 is structurally homologous to D14-3-3ε, functionally it diverges presenting characteristics of other 14-3-3s. Given the basal position of Giardia in eukaryotic evolution, this finding is consistent with the hypothesis that 14-3-3ε isoforms are ancestral to other family members.
PLoS ONE 10/2013; 8(10):e78090. · 3.53 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: In Germany and Poland, the high population density of the red fox (Vulpes vulpes) is considered a public health risk since this wild canid is one of the main reservoirs of Trichinella spp. In 2010 in Poland, a program to monitor the prevalence of Trichinella spp. in the red fox population was launched. After two years, Trichinella spp. larvae were detected in 44 (2.7%) out of 1634 foxes tested. In Germany in the period 2002-2011, Trichinella spp. larvae were in 27 foxes. The Trichinella species detected were: T. spiralis in 15 foxes from Germany (one co-infection with Trichinella britovi and one with Trichinella pseudospiralis) and in 9 foxes from Poland; T. britovi in 8 and 32 foxes from Germany and Poland, respectively; and T. pseudospiralis in 1 fox from Germany. The arctic species Trichinella nativa was detected in 3 foxes from Germany (one co-infection with Trichinella spiralis) and in 1 fox from Poland. The detection of T. nativa outside its known distribution area opens new questions on the ability of this Trichinella species to colonize temperate regions.