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.     

The fifth outbreak of trichinosis in Korea

Trichinosis is a food-borne zoonotic disease caused by the nematode, Trichinella spp., and had been reported several times in Korea. Recently, there was an additional outbreak, involving 5 patients, the findings from which are reported herein. On 30 November 2010, 8 persons ate sashimi of the meat of a wild boar. Then, 2-3 weeks later, they complained of myalgia and fever. Unfortunately, muscle biopsy was not performed, but ELISA was performed using their sera. Two people among 8 were positive for Trichinella on the 34th day post-infection (PI), and 3 patients who initially revealed negative ELISA were additionally proved to be positive for trichinosis on the 42nd day PI. Hence, the confirmed patients of trichinosis were 5 in total in the present outbreak. They were treated with albendazole and discharged uneventfully. This was the fifth outbreak of trichinosis in Korea.     

Traditional Living Habits of the Taz Tundra Population: A Paleoparasitological Study

An excavation of the Vesakoyakha II–IV and Nyamboyto I burial grounds was conducted during the 2014 field season, and soil samples from intact burials dating from the 19th and 20th centuries, respectively, were analyzed to determine interactions between parasites and host/vectors. Considering the discovery of Diphyllobothrium sp. and Taenia sp. eggs in soil samples from the pelvic region, diphyllobothriasis was the most frequent helminthic infection among the Taz Nenets. The Nyamboyto Nenets mainly consumed uncooked fish, while the Vesakoyakha Nenets had a bigger variety in food choices, including reindeer meat. Nenets children were given raw fish from early childhood. The paleoparasitological results corroborate rare ethnographic records about the consumption of uncooked reindeer cerebrum which led to beef tapeworm helminthiases. This is the first parasitological report of helminthic diseases among the Taz Nenets, and, as such, it provides insight into their subsistence activities and food patterns and broadens our understanding of their health condition.     

In Vitro Cultivation of Cymatocarpus solearis (Brachycoeliidae) Metacercariae to Obtain the Adult Stage without the Marine Turtle Definitive Host

In vitro cultivation of trematodes would assist studies on the basic biology of the parasites and their hosts. This is the first study to use the yolk of unfertilized chicken eggs as a simple and successful method of ovocultivation and the first time to obtain the adult-stage of the trematode Cymatocarpus solearis Braun, 1899 (Digenea: Brachycoeliidae). Chicken eggs were inoculated with metacercariae from the muscle of the spiny lobster, Panulirus argus (Latreille, 1804). The metacercariae were excysted and incubated for 576 hr (24 days) at 38℃ to obtain the adult stage. Eggs in utero were normal in shape and light brown color. The metacercariae developed into mature parasites that have been identified as the adult-stage found in marine turtles. The adult lobsters collected in Quintana Roo State, Mexico, showed the prevalence of 49.4% and the mean intensity of 26.0 per host (n = 87). A statistical study was performed to determine that no parasitic preference was detected for male versus female parasitized lobsters. Morphometric measurements of the adult-stage of C. solearis obtained in our study have been deposited in the National Helminths Collection of the Institute of Biology of the National Autonomous University of Mexico. This study is significant because it is the first time that a digenean of the family Brachycoeliidae has been demonstrated to develop in vitro from metacercariae into adults capable of producing eggs using the yolk of unfertilized chicken eggs. Secondly, this technique allows to obtain the adult stage of C. solearis without the presence of its marine turtle host, allows us to describe the mature parasites, and thus contribute to our understanding of the biology of C. solearis.     

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.     

Variation in reproductive output of marine turtles

Most marine turtle species are non-annual breeders and show variation in both the number of eggs laid per clutch and the number of clutches laid in a season. Large levels of inter-annual variation in the number of nesting females have been well documented in green turtle nesting populations and may be linked to environmental conditions. Other species of marine turtle exhibit less variation in nesting numbers. This inter-specific difference is thought to be linked to trophic status. To examine whether individual reproductive output is more variable in the herbivorous green turtle (Chelonia mydas Linneaeus 1758) than the carnivorous loggerhead (Caretta caretta Linneaeus 1758), we examined the nesting of both species in Cyprus over nine seasons. Green turtles showed slower annual growth rates (0.11 cm year⁻¹ curved carapace length (CCL) and 0.27 cm year⁻¹ curved carapace width (CCW)) than loggerhead turtles (0.36 cm year⁻¹ CCL, 0.51 cm year⁻¹ CCW). CCL was highly correlated to mean clutch size in both green (R²=0.51) and loggerhead turtles (R²=0.61) and maximal clutch size of green turtles (R²=0.58). Larger females did not lay a greater number of clutches or have a shorter remigration interval than smaller females of either species. On average, the size of green turtle clutches increased and that of loggerhead turtles decreased as the season progressed. Individual green turtles, however, produced more eggs per clutch through the season to a maximum in the third or fourth clutch. In loggerhead turtles, clutches 1-4 were very similar in size but the fifth clutch was 38% smaller than the first. No individuals of either species were recorded laying more than five clutches. Green turtles may not be able to achieve their maximum reproductive output with respect to clutch size throughout the season, whereas only loggerhead turtles laying five clutches (n=5) appear to become resource depleted. Green turtles nesting in years when large numbers of nests were recorded laid a greater number of clutches than females nesting in years with lower levels of nesting.     

Seroprevalence of Encephalitozoon cuniculi and Toxoplasma gondii in domestic rabbits (Oryctolagus cuniculus) in China

The breeding of domestic rabbits (Oryctolagus cuniculus) for human consumption has a long tradition in China. Infections that can affect the production of meat or even be transmitted from animals to humans are important to monitor, especially for public health reasons as well as for their impact on animal health. Thus, a total of 1,132 domestic rabbit sera from 4 regions in China were collected for serological screening for Encephalitozoon cuniculi and for Toxoplasma gondii by ELISA and modified agglutination test (MAT), respectively. Antibodies to E. cuniculi were detected in 248/1,132 (21.9%) sera tested while antibodies against T. gondii revealed a seroprevalence of 51/1,132 (4.5%). We believe that the present results are of epidemiological implications and public health importance due to the acknowledged susceptibility of humans to E. cuniculi and T. gondii infections. Therefore, routine screening tests of domestic rabbits are proposed considering the zoonotic potential of these parasites.     

To Eat or not to Eat an Endangered Species: Views of Local Residents and Physicians on the Safety of Sea Turtle Consumption in Northwestern Mexico

Sea turtles have historically been an important food resource for many coastal inhabitants of Mexico. Today, the consumption of sea turtle meat and eggs continues in northwestern Mexico despite well-documented legal protection and market conditions providing easier access to other more reliable protein sources. Although there is growing evidence that consuming sea turtles may be harmful to human health due to biotoxins, environmental contaminants, viruses, parasites, and bacteria, many at-risk individuals, trusted information sources, and risk communicators may be unaware of this information. Therefore, we interviewed 134 residents and 37 physicians in a region with high rates of sea turtle consumption to: (1) examine their knowledge and perceptions concerning these risks, as a function of sex, age, occupation, education and location; (2) document the occurrence of illness resulting from consumption; and (3) identify information needs for effective risk communication. We found that 32% of physicians reported having treated patients who were sickened from sea turtle consumption. Although physicians believed sea turtles were an unhealthy food source, they were largely unaware of specific health hazards found in regional sea turtles, regardless of location. By contrast, residents believed that sea turtles were a healthy food source, regardless of sex, age, occupation, and education, and they were largely unaware of specific health hazards found in regional sea turtles, regardless of age, occupation, and education. Although most residents indicated that they would cease consumption if their physician told them it was unhealthy, women were significantly more likely to do so than men. These results suggest that residents may lack the necessary knowledge to make informed dietary decisions and physicians do not have enough accurate information to effectively communicate risks with their patients.     

Antibiogram profiling and pathogenic status of Aeromonas species recovered from Chicken

Poultry meat and its products are widely consumed by humans globally, however, Aeromonas infections in poultry have been reported in different parts of the world with devastating effects. This study was carried out to assess the antibiogram and biofilm forming potential of Aeromonas isolated from chicken fecal samples. Aeromonas isolates were screened for antibiotic susceptibility using antibiotics disk and biofilm producing potentials on abiotic surfaces. Nineteen isolates recovered from chicken feces were 100% sensitive to ciprofloxacin, gentamicin and the tetracyclines. About 53% of Aeromonas isolates were resistant to erythromycin and 47% resistant to streptomycin. Eight isolates (42.1%) were found to be moderate producers of biofilm, 31.6% (6/19) were weak producers of biofilm, 10.5% (2/19) were non biofilm producers while 15.8% (3/19) were strong producers. The present investigation shows a prevalence of potentially pathogenic Aeromonas strains in chicken feces, suggesting potential group at risk for Aeromonas infection which could be dissemination to other animals or humans with close contact and the wider community.     

Proteome changes underpin improved meat quality and yield of chickens (Gallus gallus) fed the probiotic Enterococcus faecium

Background

Supplementation of broiler chicken diets with probiotics may improve carcass characteristics and meat quality. However, the underlying molecular mechanism remains unclear. In the present study, 2D-DIGE-based proteomics was employed to investigate the proteome changes associated with improved carcass traits and meat quality of Arbor Acres broilers (Gallus gallus) fed the probiotic Enterococcus faecium.

Results

The probiotic significantly increased meat colour, water holding capacity and pH of pectoral muscle but decreased abdominal fat content. These meat quality changes were related to the altered abundance of 22 proteins in the pectoral muscle following E. faecium feeding. Of these, 17 proteins have central roles in regulating meat quality due to their biological interaction network. Altered cytoskeletal and chaperon protein expression also contribute to improved water holding capacity and colour of meat, which suggests that upregulation of chaperon proteins maintains cell integrity and prevents moisture loss by enhancing folding and recovery of the membrane and cytoskeletal proteins. The down-regulation of β-enolase and pyruvate kinase muscle isozymes suggests roles in increasing the pH of meat by decreasing the production of lactic acid. The validity of the proteomics results was further confirmed by qPCR.

Conclusions

This study reveals that improved meat quality of broilers fed probiotics is triggered by proteome alterations (especially the glycolytic proteins), and provides a new insight into the mechanism by which probiotics improve poultry production.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-1167) contains supplementary material, which is available to authorized users.     

Therapeutic Potential of Myrrh and Ivermectin against Experimental Trichinella spiralis Infection in Mice

Trichinosis is a parasitic zoonosis caused by the nematode Trichinella spiralis. Anthelmintics are used to eliminate intestinal adults as well as tissue-migrating and encysted larvae. This study aimed to investigate the effects of ivermectin and myrrh obtained from the aloe-gum resin of Commiphora molmol on experimental trichinosis. Ninety albino mice were orally infected with 300 T. spiralis larvae. Drugs were tested against adult worms at day 0 and day 5 and against encysted larvae on day 15 and day 35 post-infection (PI). Mature worms and encysted larvae were counted in addition to histopathological examination of muscle specimens. Serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), total protein, albumin, globulin, urea, and creatinine values were estimated. Significant reductions in mean worm numbers were detected in ivermectin treated mice at day 0 and day 5 PI achieving efficacies of 98.5% and 80.0%, while efficacies of myrrh in treated mice were 80.7% and 51.5%, respectively. At days 15 and 35 post-infection, ivermectin induced significant reduction in encysted larval counts achieving efficacies of 76.5% and 54.0%, respectively, while myrrh efficacies were 76.6% and 35.0%, respectively. AST, ALT, urea, and creatinine levels were reduced, while total proteins were increased in response to both treatments compared to their values in the infected non-treated mice. Ivermectin use for controlling T. spiralis could be continued. Myrrh was effective and could be a promising drug against the Egyptian strains of T. spiralis with results nearly comparable to ivermectin.     

An Outbreak of Trichinosis with Molecular Identification of Trichinella sp. in Vietnam

The 5th outbreak of trichinosis occurred in a mountainous area of North Vietnam in 2012, involving 24 patients among 27 people who consumed raw pork together. Six of these patients visited several hospitals in Hanoi for treatment. Similar clinical symptoms appeared in these patients within 5-8 days after eating infected raw pork, which consisted of fever, muscle pain, difficult moving, edema, difficult swallowing, and difficult breathing. ELISA revealed all (6/6) positive reactions against Trichinella spiralis antigen and all cases showed positive biopsy results for Trichinella sp. larvae in the muscle. The larvae detected in the patients were identified as T. spiralis (Vietnamese strain) by the molecular analysis of the mitochondrial cytochrome c oxidase subunit III (cox3) gene.     

The dual function of the lung in chelonian sea turtles: buoyancy control and oxygen storage

Chelonian sea turtles use their lung as a buoyancy organ and as the major oxygen store when diving, and hence, buoyancy regulation and oxygen consumption can be expected to interact. The buoyancy of seven juvenile loggerhead turtles, Caretta caretta, was determined by measuring directly their underwater weight (M_uw) while they were resting on a freely suspended weighing platform at a depth of 80 cm. Underwater weight was recorded continuously for 2 days for each turtle, followed by another 2 days measurements during which the turtles carried lead weights attached to their carapace, and finally, a last day of measurement after the weights had been removed. Total duration of resting dives (t_r), buoyancy (F_B), total resulting force acting on the resting turtle (F_res) and body volumes were derived from the M_uw data. Turtles were slightly negatively buoyant when resting (F_res=−0.2943 to −0.981 N kg⁻¹) and M_uw increased significantly throughout each apnoeic period, meaning that the turtles progressively lost buoyancy. Pulmonary gas loss was calculated from the rate at which buoyancy decreased, which was significantly slower during the first half of the dive than during the second half of the dive. Resting oxygen consumption rates (V_O2) were calculated from these data assuming that the pulmonary gas loss represents oxygen consumption from the lung. The V_O2 obtained in this way (1.69-4.86 ml O₂ min⁻¹) corresponded well with other published and V_O2 measured previously on loggerhead turtles in the same facility. Using oxygen from the lung affects buoyancy and may have impacts on the diving behaviour.     

First fossil gravid turtle provides insight into the evolution of reproductive traits in turtles

Here we report on the first discovery of shelled eggs inside the body cavity of a fossil turtle and on an isolated egg clutch, both referable to the Cretaceous turtle Adocus. These discoveries provide a unique opportunity to gain insight into the reproductive traits of an extinct turtle and to understand the evolution of such traits among living turtles. The gravid adult and egg clutch indicate that Adocus laid large clutches of rigid-shelled spherical eggs and established their nests near rivers, traits that are shared by its closest living relatives, the soft-shelled turtles. Adocus eggshell, however, was probably more rigid than that of living turtles, based on its great thickness and structure, features that may represent unique adaptations to intense predation or to arid nest environments. In light of the reproductive traits observed in Adocus, the distribution of reproductive traits among turtles reveals that large clutches of rigid-shelled eggs are primitive for hidden-necked turtles (cryptodirans) and that spherical eggs may have evolved independently within this group.     

An innovative survey underlining the significant level of contamination by Toxoplasma gondii of ovine meat consumed in France

Consumption of sheep meat presents a risk of human contamination by Toxoplasma gondii. A nationwide study was conducted in France to evaluate the prevalence of Toxoplasma in fresh ovine meat. A sampling procedure was established to guarantee the representativity of consumption. As is the case for meat consumed, half of the samples were from France and half were imported from other countries. Animals were selected according to their age, as lamb (<12 months) represents 90% of the meat consumed. Available data for French samples allowed the selection of 16 districts distributed in seven areas according to their density of production. Diaphragms and hearts from 433 sheep were collected. Diaphragms were collected from 398 imported carcasses. Fluids from hearts and diaphragms were tested serologically. All hearts were bioassayed in mice and parasite isolates were genotyped using PCR-restriction fragment length polymorphism and microsatellite markers. Prevalence estimates were calculated, taking into account uneven distribution of production and age. For French meat, the effect of area, age and their interactions was evaluated. The overall estimate of Toxoplasma seroprevalence was 17.7% (11.6-31.5%) for lambs and 89% (73.5-100%) for adults (P < 0.0001). No significant difference was observed between imported and French meat. In France, seroprevalence in lambs showed an increasing North-western to Southern gradient. The proportion of French carcasses carrying live parasites according to bioassay results was estimated at 5.4% (3-7.5%) (45 genotype II; one genotype III). This study offers an accurate drawing of the toxoplasmosis pattern amongst sheep consumed in France and a model for a zoonosis hazard control survey.     

Expression patterns of peroxisome proliferator-activated receptor gamma 1 versus gamma 2, and their association with intramuscular fat in goat tissues

Intramuscular fat (IMF) shortage causes the lack of juiciness and tenderness of goat meat, while peroxisome proliferator-activated receptor gamma 1 (PPARγ1) and gamma 2 (PPARγ2) play key roles in lipid metabolism. Nevertheless, their expression patterns and the relationship with IMF have been poorly exposed. Using quantitative polymerase chain reaction (qPCR), classical Soxhlet extraction, and in situ hybridization, we demonstrated that among 13 goat tissues, expression of PPARγ1 was dramatically higher than that of PPARγ2 except for lung. We further demonstrated the expression patterns of PPARγ1 and PPARγ2 and their negative association with intramuscular fat content in three goat muscles with kids growing. Meanwhile, PPARγ expression was located in the connective tissues. These results suggest that PPARγ1 is rather active for most tissues of goat, and closely related with the muscular fat metabolism during early postnatal life, but a more direct proof remains to be provided.     

Seropositivity of Toxoplasmosis in Pregnant Women by ELISA at Minia University Hospital,Egypt

Toxoplasmosis is considered as an important risk factor for bad obstetric history (BOH) and one of the major causes of congenitally acquired infections. The present study aimed to estimate the seropositivity of T. gondii infection and associated risk factors among the attendees of high risk pregnancy and low risk antenatal care clinic of Minia Maternity and Pediatric University Hospital, Minia, Egypt. The study was carried out from April 2013 to April 2014 through 2 phases, the first phase was case-control study, and the second phase was follow-up with intervention. A total of 120 high risk pregnant and 120 normal pregnant females were submitted to clinical examinations, serological screening for anti-Toxoplasma IgM and IgG antibodies by ELISA, and an interview questionnaire. Seropositive cases were subjected to spiramycin course treatment. The results showed that the seroprevalence of toxoplasmosis in high-risk pregnancy group was 50.8%, which was significantly different from that of normal pregnancy group (P<0.05). Analysis of seropositive women in relation to BOH showed that abortion was the commonest form of the pregnancy wastage (56.5%). The high prevalence of T. gondii seropositive cases was observed in the age group of 21-30 years. Post-delivery adverse outcome was observed in 80.3% of high-risk pregnancy group compared to 20% of normal pregnancy group. There was a statistically significant relationship between seropositivity and living in rural area, low socioeconomic level, and undercooked meat consumption (P<0.05). Serological screening for anti-Toxoplasma antibodies should be routine tests especially among high-risk pregnant women.     

Toxoplasma gondii B1 Gene Detection in Feces of Stray Cats around Seoul,Korea and Genotype Analysis of Two Laboratory-Passaged Isolates

The increasing prevalence of Toxoplasma gondii infection in the human population in the Republic of Korea (= Korea) is due to various reasons such as an increase in meat consumption. However, the importance of cats in transmitting T. gondii infection through oocysts to humans has seldom been assessed. A total of 300 fecal samples of stray cats captured around Seoul from June to August 2013 were examined for T. gondii B1 gene (indicating the presence of oocysts) using nested-PCR. Fourteen (4.7%) of 300 cats examined were positive for B1 gene. Female cats (7.5%) showed a higher prevalence than male cats (1.4%). Cats younger than 3 months (5.5%) showed a higher prevalence than cats (1.5%) older than 3 months. For laboratory passage of the positive samples, the fecal suspension (0.2 ml) of B1 gene positive cats was orally inoculated into experimental mice. Brain tissues of the mice were obtained after 40 days and examined for the presence of tissue cysts. Two isolates were successfully passaged (designated KNIH-1 and KNIH-2) and were molecularly analyzed using the SAG5D and SAG5E gene sequences. The SAG5D and SAG5E gene sequences showed high homologies with the ME49 strain (less virulent strain). The results indicated the importance of stray cats in transmitting T. gondii to humans in Korea, as revealed by detection of B1 gene in fecal samples. T. gondii isolates from cats were successfully passaged in the laboratory for the first time in Korea.     

Complete amino acid sequence of three reptile lysozymes

To study the structure and function of reptile lysozymes, we have reported their purification, and in this study we have established the amino acid sequence of three egg white lysozymes in soft-shelled turtle eggs (SSTL A and SSTL B from Trionyx sinensis, ASTL from Amyda cartilaginea) by using the rapid peptide mapping method. The established amino acid sequence of SSTL A, SSTL B, and ASTL showed substitutions of 43, 42, and 44 residues respectively when compared with the HEWL (hen egg white lysozyme) sequence. In these reptile lysozymes, SSTL A had one substitution compared with SSTL B (Gly126Asp) and had an N-terminal extra Gly and 11 substitutions compared with ASTL. SSTL B had an N-terminal extra Gly and 10 residues different from ASTL. The sequence of SSTL B was identical to soft-shelled turtle lysozyme from STL (Trionyx sinensis japonicus). The Ile residue at position 93 of ASTL is the first report in all C-type lysozymes. Furthermore, amino acid substitutions (Phe34His, Arg45Tyr, Thr47Arg, and Arg114Tyr) were also found at subsites E and F when compared with HEWL. The time course using N-acetylglucosamine pentamer as a substrate exhibited a reduction of the rate constant of glycosidic cleavage and increase of binding free energy for subsites E and F, which proved the contribution for amino acids mentioned above for substrate binding at subsites E and F. Interestingly, the variable binding free energy values occurred on ASTL, may be contributed from substitutions at outside of subsites E and F.