Enzyme-linked immunosorbent assay for detection of Trichinella spiralis antibodies and the surveillance of selected pig breeding farms in the Republic of Korea

Trichinellosis is a parasitic zoonosis of public health importance. It is caused by Trichinella spiralis which has a wide host range including humans. In the present communication, the ELISA technique was employed on a total of 803 blood samples from 7 selected pig breeding farms in 1996 for diagnosis and surveillance of trichinellosis. Out of the entire 803 samples, nine were found to be suspected while one was positive by ELISA. But western blot analyses employed for further confirmation have shown that all of 10 samples did not react to larval excretory-secretory product antigens. These results indicate that pig breeding farms included in the present study are free from trichinellosis. However, it does not mean Korea is free from trichinellosis since human trichinellosis has recently been reported. The necessity of continued surveillance for trichinellosis in both pigs and wild animals was discussed.     

Molecular identification of a Trichinella isolate from a naturally infected pig in Tibet, China

The first human case with trichinellosis was reported in 1964 in Tibet, China. However, up to the present, the etiological agent of trichinellosis has been unclear. The aim of this study was to identify a Tibet Trichinella isolate at a species level by PCR-based methods. Multiplex PCR revealed amplicon of the expected size (173 bp) for Trichinella spiralis in assays containing larval DNA from Tibet Trichinella isolate from a naturally infected pig. The Tibet Trichinella isolate was also identified by PCR amplification of the 5S ribosomal DNA intergenic spacer region (5S ISR) and mitochondrial large-subunit ribosomal RNA (mt-lsrDNA) gene sequences. The results showed that 2 DNA fragments (749 bp and 445 bp) of the Tibet Trichinella isolate were identical to that of the reference isolates of T. spiralis. The Tibet Trichinella isolate might be classifiable to T. spiralis. This is the first report on T. spiralis in southwestern China.     

Social,political, and economic factors responsible for the reemergence of trichinellosis in Serbia: a case study

Over the past decade, eastern Europe has experienced a resurgence of trichinellosis. A recent outbreak in Serbia, Yugoslavia, from December 2001 to January 2002, involving 309 people, revealed many of the causes for this reemergence. Epidemiological investigations indicate that the immediate cause of the recent outbreak was the consumption of smoked sausages produced by a small slaughterhouse or meat processor. However, failure of in-house meat inspection procedures and quality assurance as well as oversight by official veterinary control were also responsible. Further analysis of this breakdown in the food safety net revealed additional general factors that have yielded a seriously deficient veterinary control system, and these are factors that are relevant to the problems experienced throughout eastern Europe and other regions. The recent civil war that led to the breakup of the former Federation of Yugoslavia resulted in severe economic and demographic changes, including high inflation and external economic sanctions. This led to (1) the loss of large numbers of experienced veterinary control officers and their replacement with inexperienced personnel, (2) a change in the swine industry with reduction in the number of large establishments with in-house inspection and replacement with more than 1,000 small abattoirs, too small to afford full-time in-house inspection, and (3) an increase in smallholder pig farming with reduced government oversight to ensure high standards in pig-rearing practices (infection risk management). The consequences of these events have been a 300% increase in Serbian pig infection and a concomittant large increase in human outbreaks. Before 1990, swine trichinellosis in Serbia was confined to 4 small districts, but today about one third of the Republic is considered endemic for trichinellosis. The reemergence of trichinellosis in Serbia illustrates the ability of this zoonosis to "leak" through a poorly maintained food safety barrier and the vulnerability of effective veterinary control to national and international events.     

Seroprevalence of Trichinella sp. in Wild Boars (Sus scrofa) from Yanggu-gun,Gangwon-do,Korea

A total 7 outbreaks of trichinellosis have occurred in Korea, mostly as a result of consumption of raw wild boar (Sus scrofa) meat. Since only 1 serological survey on wild boars had yet been performed in Korea, the present study aimed to estimate the prevalence of trichinellosis in wild boars and some species of rodents by artificial digestion and serological examinations in Yanggu-gun, Gangwon-do, the endemic area of trichinellosis. Both the wild boar and rodent muscle samples revealed no Trichinella larvae by direct examination and artificial digestion method. However, serological examinations revealed that 4 wild boar sera samples out of 118 (3.4%) were positive to Trichinella antigen. Although the recovery of Trichinella larvae ended in a failure, it is proved for the first time that the sylvatic cycle of Trichinella has been maintained in wild boars of Gangwon-do, Korea.     

Immunoelectrotransfer blot assay in acute and chronic human trichinellosis

An immunoelectrotransfer blot assay (IETB) using excretory secretory products of muscle larvae of Trichinella spiralis (ML-ESP) and the avidin biotin system was developed in order to characterize reactivity against ML-ESP in sera from patients with acute and chronic trichinellosis. A complete pattern of up to 13 bands was developed by sera from individuals with trichinellosis where doublets, triplets, or single bands were shown to have molecular weights of roughly 66, 55, 45, 36, 29, 24, and 14 kDa. The bands at approximately 55, 36, 29, and 14 kDa proved specific for T. spiralis. The band at approximately 55 kDa was present in all trichinellosis sera, whereas the approximately 14-kDa band was present in only a small percentage of sera. The development of approximately 36- and 29-kDa bands suggests a modulation of the reactivity against ML-ESP over time. IETB proved more sensitive for the population of chronic trichinellosis under study than a conventional diagnostic enzyme-linked immunosorbent assay, allowing negative or borderline serum samples to be determined. Thus, this technique, when applied for human trichinellosis surveillance, should provide a useful tool in endemic areas.     

Differential diagnosis of schistosomiasis mekongi and trichinellosis in human.

An indirect (plate) ELISA and, a more convenient version, a dot-blot (membrane) ELISA have been developed using haemocyanin of a mollusk, Megathura crenulata, i.e. keyhole limpet haemocyanin (KLH) and purified, specific antigen of Trichinella spiralis (APTsAg) obtained from a monoclonal antibody-affinity column chromatography, for differential diagnosis of schistosomiasis mekongi and trichinellosis. Serum samples of patients with parasitologically confirmed trichinellosis were reactive to both antigens in both versions of ELISA while sera of patients with schistosomiasis mekongi were positive only to the KLH. Both ELISA were negative when used to test sera of normal controls and patients with gnathostomiasis, paragonimiasis and opisthorchiasis.     

Importance of the wild boar in the epidemiology of wild trichinellosis in Piedmont and Liguria

Muscle samples from 1,508 wild boars (Sus scrofa) harvested in Piedmont and Liguria between 1987 and 1990 were examined for larvae of Trichinella sp. by trichinelloscopy and peptic digestion. No wild boar was infected, though trichinellosis was present in foxes (Vulpes vulpes) from the same areas (14 positives out of 608) with the highest prevalence in the mountain range (25.0%). Infection was also not found in 40 stone martens, 5 badgers and 3 stoats. Five vulpine isolates were identified as belonging to T3 zymodeme. These results show that free-ranging wild boars, although being a source of trichinellosis infection for man in Italy (9.2% of the 584 cases diagnosed since 1961), have no significance in maintaining the sylvatic cycle of this parasite in the study area. The data agree with the low infectivity of Italian strains of fox origin for swine seen in the experimental infections carried out so far.     

Diallyl sulfide--a flavour component from garlic (Allium sativum) attenuates lipid peroxidation in mice infected with Trichinella spiralis.

We investigated the antioxidant capacity of diallyl sulfide (DAS) in the course of experimental trichinellosis in C3H mice. For this purpose, the mice were orally infected with either Trichinella spiralis larvae (300 larvae/mouse) or treated per os with normal saline (control), and 12 hrs later, they were treated per os with DAS (200 mg/kg b.w) daily for 5 or 20 days. On day 5 and 35 of the post-infection period (5 and 35 dpi), thiobarbituric-acid reactive substances (TBARS) in the small intestinal mucosa and diaphragm samples and the total antioxidant status (TAS) of blood were determined. Trichinella spiralis increased TBARS and decreased TAS in the intestinal phase of invasion. However, in the muscular phase mice, neither TBARS nor TAS was found to be different from those of the control. Diallyl sulfide has been shown to decrease TBARS and the agent did not have any effect(s) on the total antioxidant status of blood in Trichinella-infected mice. The results suggest that diallyl sulfide may be an effective antioxidant candidate and may therefore play a significant role in the defense against lipid peroxidation in trichinellosis.     

An Outbreak of Trichinellosis by Consumption of Raw Soft-Shelled Turtle Meat in Korea

Trichinellosis transmission to humans via the consumption of reptile meat is rare worldwide. In Korea, however, 2 such outbreaks, possibly via consumption of soft-shelled turtle meat, have occurred in 2 successive years. In 17 August 2014, 6 patients were admitted to Wonju Severance Christian Hospital complaining of myalgia, fever, and headache. Eosinophilia was the indication of the initial laboratory results, and they were eventually diagnosed as trichinellosis by ELISA. All of the patients worked at the same company and had eaten raw soft-shelled turtle meat at a company dinner 10 days prior to their admission. They were treated with albendazole for 2 weeks, upon which all of their symptoms disappeared. This is the 8th report on human trichinellosis in Korea, and the second implicating raw soft-shelled turtle meat.     

Inhibition of leukocyte function by serum from patients with trichinellosis

Modification of leukocytic function has been reported in only a few human parasitic diseases. In this study we evaluated the effects of the sera from patients infected with Trichinella on chemotactic and phagocytic responses in leukocytes. Leukocyte chemotaxis was tested by the agarose method and phagocytosis by the technique of Yamamura, modified for Saccharomyces cerevisiae. Sera were acquired from patients during a trichinellosis outbreak that occurred in northern Italy in 1986. The parasite was isolated from 1 patient and isoenzymatically typed as Trichinella sp. 3, a new taxon, previously considered Trichinella nelsoni. The results indicated that sera from Trichinella-infected humans inhibited both chemotaxis and phagocytic responses in leukocytes. These findings suggest the existence of serum factor(s) in trichinellosis patients that modify host leukocytic functions. The source and nature of active serum components and the mechanism by which they modulate leukocyte function remain to be clarified.     

Comparative efficacy of antigen and antibody detection tests for human trichinellosis

Sera collected from patients with suspected or confirmed exposure to Trichinella spiralis were tested for circulating parasite antigens and antiparasite antibodies. Using an immunoradiometric assay, excretory--secretory antigens from muscle-stage larvae of T. spiralis were detected in the sera of 47% of 62 patients with clinical trichinellosis and 13% of 39 patients without clinical signs but suspected of exposure to infected meat. In comparison, antibodies were detected using an indirect immunofluorescent test in the circulation of 100% of the 62 patients with clinical trichinellosis and 46% of the 39 patients with suspected exposure. The presence of antibodies specific to excretory-secretory products of T. spiralis muscle larvae was confirmed in the majority of the samples tested by a monoclonal antibody-based competitive inhibition assay. These results indicate that antibody detection is a more sensitive diagnostic method for human trichinellosis, but that antigen detection might be a useful confirmatory test because it is a direct demonstration of parasite products in the circulation.     

Trichinellosis: the zoonosis that won't go quietly

Trichinellosis, is normally not included among those regarded as emerging zoonoses because it has been a public health threat for more than 150 years. However, its dramatic re-emergence in many areas around the world over the past 10-20 years, inspite of a century of veterinary public health efforts to control and eradicate it, justifies it being included in this group. The reasons for this re-emergence are diverse, and include human pertubation and manipulation of ecosystems, war and political turmoil, rapidly changing food distribution and marketing systems, and even, surprisingly, rising affluence in developing countries. These influences, and their impact on the epidemiology of both domestic and sylvatic trichinellosis, are discussed, along with recommendations for confronting this altered status as a public health threat.     

Trichinella spiralis in an agricultural ecosystem. II. Evidence for natural transmission of Trichinella spiralis spiralis from domestic swine to wildlife

Epidemiological investigations of an outbreak of trichinellosis were carried out in a domestic swine herd and it was established that the parasite also occurred in rats, and in skunks, opossums, and raccoons. Because considerable uncertainty exists regarding the role of sylvatic trichinellosis as a reservoir for the synanthropic cycle, studies were conducted to determine the genetic nature of the various isolates from this ecosystem. Pig infectivity trials, isoenzyme analyses, and repetitive DNA sequence analyses were performed. The results showed that all isolates from the farm environs were genetically similar and that they are related to Trichinella spiralis isolated from domestic pigs. The implication of these findings, in contrast to studies on isolates from wildlife elsewhere, is that this parasite is transmitted from domestic swine to sylvatic hosts and that any control or eradication efforts must take into account the potential for reinfection of hogs from wild animals.     

Fish-borne parasitic zoonoses: status and issues

The fish-borne parasitic zoonoses have been limited for the most part to populations living in low- and middle-income countries, but the geographical limits and populations at risk are expanding because of growing international markets, improved transportation systems, and demographic changes such as population movements. While many in developed countries will recognize meat-borne zoonoses such as trichinellosis and cysticercosis, far fewer are acquainted with the fish-borne parasitic zoonoses which are mostly helminthic diseases caused by trematodes, cestodes and nematodes. Yet these zoonoses are responsible for large numbers of human infections around the world. The list of potential fish-borne parasitic zoonoses is quite large. However, in this review, emphasis has been placed on liver fluke diseases such as clonorchiasis, opisthorchiasis and metorchiasis, as well as on intestinal trematodiasis (the heterophyids and echinostomes), anisakiasis (due to Anisakis simplex larvae), and diphyllobothriasis. The life cycles, distributions, epidemiology, clinical aspects, and, importantly, the research needed for improved risk assessments, clinical management and prevention and control of these important parasitic diseases are reviewed.     

Human trichinellosis in Hungary from 1965 to 2009

Human trichinellosis was first documented in Hungary in 1891 and then there were an increased number of reports up to 1964 when the most severe outbreak occurred. After that, no information was available on the international literature on human trichinellosis which occurred from 1965 up to the present years. The aim of this study was to collect all the data available in Hungarian official data-sources on human trichinellosis which occurred from 1965 up to 2009 in Hungary. Furthermore, a comparative analysis was performed on the different serological tests used along the 45 years of investigation. In the period in question, 573 infections were documented in Hungary. Of them, 57 occurred in the years 1965-69, 130 in 1970-79, 302 in 1980-89, 27 in 1990-99, and 57 in 200009. The most common sources of infection were pork from backyard pigs and hunted wild boars. Sporadic cases and small family outbreaks marked the last ten years. The comparison of serological tests shows that the ELISA is a good test for the first screening, but ELISA-positive serum samples should be confirmed by western blot except for clinically clear-cut cases.     

Frequency and severity of musculoskeletal symptoms in humans during an outbreak of trichinellosis caused by Trichinella britovi

Musculoskeletal symptoms such as myalgia are well-known features in the course of trichinellosis; however, the characteristics of musculoskeletal findings have been described in detail in only 1 study. The present study was aimed to determine the joint and muscle symptoms in subjects diagnosed with acute trichinellosis at our rheumatology unit during a Trichinella britovi outbreak that occurred in Izmir, Turkey, in 2004. In total, 98 patients (55 females, 43 males; mean age 32.3 +/- 10.9 yr) were included in the study. A detailed history and full musculoskeletal examination were obtained in each patient. A self-administered questionnaire developed for recording the musculoskeletal symptoms was completed monthly until all the symptoms were resolved. Pain at the joints, restriction of movements (in shoulders, elbows, wrists, knees, ankles, and temporomandibular joints), myalgia, and muscle weakness (neck and shoulder girdle, muscles of the upper and forearm, back, thigh, and calf muscles) were assessed in every patient. Eosinophil counts, serum levels of creatine kinase, and lactate dehydrogenase also were analyzed. The most frequent musculoskeletal symptoms were muscle pain (86 cases [87.8%]), joint pain (83 [84.7%]), subjective muscle weakness (75 [76.5%]), and restriction of joint movements (63 [64.3%]). Calves, upper arm, neck and shoulder girdle, and forearms were the most affected muscle groups. Muscle pain was reported more frequently in the upper than in the lower extremities and during activity. The most frequent painful joints were shoulders, knees, wrists, and ankles. Upper extremity joints were affected more frequently than the lower extremity joints (77.6 vs. 70.4%). Joint pain occurred more frequently at rest. Both muscle weakness and restriction of joint movements were reported in and around the most frequently affected regions. No evidence of arthritis and objective muscle weakness was noted on physical examination in any patient. Musculoskeletal symptoms in the course of T. britovi infection are frequent but with an excellent prognosis. Joint pain in people suffering from acute trichinellosis may occur more frequently than reported previously.     

Application of GIS in epizootiological surveillance of swine trichinellosis in one endemic district in Serbia

Application of new tools for epizootiological investigations in veterinary medicine, such as Geographical Information Systems (GIS), offers a new approach and possibilities for the eradication or control of infectious diseases. GIS is particularly useful for research conducted in small areas strongly impacted by man. Trichinellosis is a world-wide zoonosis, which is endemic in some European countries, Balkan district and Serbia in particular. There are very few data on GIS application in the field of trichinellosis. We here present the application of GIS for mapping Trichinella spp. occurrence and spatial and temporal patterns of Trichinello infection in one endemic district in Serbia. Settlements with trichinellosis were marked and particular points of interest were designated. Data on prevalence of Trichinella infection in domestic swine accompanied by location of foci indicated the existence of disease geographical stationarity. This first report on GIS application in Serbia will facilitate trichinellosis surveillance and monitoring of Trichinella spp. circulation among domestic pigs, and populations of synanthropic and sylvatic animals.     

Patterns and Risks of Trichinella Infection in Humans and Pigs in Northern Laos

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.