Effectiveness of health education in improving knowledge, attitude and practice related to foodborne zoonotic trematodes in Vietnam, with a particular focus on Clonorchis sinensis.

OBJECTIVE: Foodborne zoonotic trematodes (FZT), which infect the liver, lungs and intestines of humans, are an emerging public health concern in tropical countries including Vietnam. In northern Vietnam, Clonorchis sinensis is recognised as the most important species of the FZT. Up to now, small-scale studies conducted in the country have indicated that health education could improve participants' knowledge and practices related to clonorchiasis, however strong evidence is still lacking. We conducted an intervention trial between 2020 and 2021 in four communes in Yen Bai and Thanh Hoa province, aiming to evaluate the impact of an extended educational package on knowledge, attitude and practices related to clonorchiasis, as well as on knowledge on the large liver fluke and minute intestinal flukes. METHODS: To this end, baseline and post-intervention questionnaires were conducted. Generalised estimating equations models were run to analyse the impact of the intervention on knowledge, attitudes and practices over time in the intervention versus control group. Finally, 172 people in the intervention group and 162 in the control group were included for analysis. RESULTS: Results demonstrated that the intervention significantly improved clonorchiasis knowledge, attitudes and practices, with higher odds for a correct knowledge response (odds ratio (OR) = 2.80, 95% confidence interval (CI) = 1.84-4.27, p-value < 0.001), increased average mean attitude score (mean = 0.363, 95%CI = 0.182-0.544, p-value < 0.001), and a reduced odds of consuming raw fish (OR = 0.15, 95%CI = 0.06-0.40, p = 0.002) in the intervention group compared to the control group post-intervention versus baseline. Additionally, participants in the intervention group exhibited enhanced basic knowledge of the large liver fluke and minute intestinal flukes post-intervention. CONCLUSIONS: These findings demonstrate the potential of health education programs in preventing clonorchiasis in endemic areas, emphasising the importance of continued health education as a critical component of integrated control programs for clonorchiasis.

Environmental influence on abundance and infection patterns of snail intermediate hosts of liver and intestinal flukes in North and Central Vietnam.

Liver and intestinal flukes (LIF) are important groups of foodborne zoonotic trematodes (FZTs) in Southeast Asia, including Vietnam. Their complex life cycles require specific freshwater snail species as the obligatory first intermediate hosts. In 2019, we conducted a longitudinal study in Yen Bai and Thanh Hoa provinces in North and Central Vietnam, respectively, to investigate the diversity of LIF and their infection prevalence in relation to snail host abundance and environmental factors. Using a combination of morphological and molecular identification techniques, we identified 10 LIF species infecting 11 snail host species. We observed significant seasonal variation in the mean abundance of several snail host species, with the majority of snails collected during the spring. We also detected seasonal changes in LIF species composition, with the highest species richness reported in the spring. Clonorchis sinensis and Fasciola gigantica, two medically important human liver flukes in Asia, were found only in the spring in Yen Bai. Our study revealed that not all snail host species have the same probability of becoming infected, and we recorded seasonal variations in the prevalence of LIF infection in different snail species in relation to water parameters.

Incidence of fish-borne trematode infections and associated factors: results from a cohort study in highly endemic communities in northern Vietnam.

Fish-borne trematodes (FiBT) are an important group of zoonotic parasites negatively affecting human health, mainly in Asia. Most studies on FiBT have applied a cross-sectional design, which provides weaker evidence on potential risk factors for transmission than a cohort study. This cohort study aimed to estimate the incidence and identify associated risk factors for FiBT infection in Vietnam. Between April 2018 and May 2019, two communes in Yen Bai province, a highly endemic area for FiBT, were visited for sampling. Participants with a negative stool result for FiBT at baseline, were invited for follow-up and data collection, at months 4, 9, and 13. Stools were examined using Kato-Katz and formalin-ethyl acetate concentration techniques to detect FiBT eggs, whereas a questionnaire was used for interviewing participants to determine the risk factors for FiBT infection during each follow-up period. The incidence risk and the incidence rate were calculated, and univariate and multivariable models were run to identify the risk factors for FiBT. A total of 194 people, negative for FiBT eggs at the baseline survey, were invited to participate in the study, and 111 people agreed to enroll in the follow-up. The incidence risk at months 4, 9, and 13 was 9.0%, 6.4%, and 5.1%, respectively. We finally used data from 95 participants for the risk factor analysis, excluding 16 people lost for the follow-up. Overall, 20 people became infected with FiBT (IR = 21.1%). The incidence rate of FiBT infection was 21.4/100 person-year. In the univariate analysis, consumption of raw fish was the main risk factor (RR = 4.59, 95%CI = 1.95-10.82), followed by being male (RR = 3.41, 95%CI = 1.56-7.45) and drinking alcohol (RR = 3.25, 95%CI = 1.49-7.11). In the multivariable analysis, only consumption of raw-fish dishes was significantly associated with FiBT infection. The people who consumed raw fish were 3.44 (95%CI = 1.11-10.70) times more at risk of infection with FiBT as compared to individuals who did not consume raw fish. It can be concluded that the FiBT incidence is high in the study area. More awareness campaigns are needed to stop eating raw fish in these areas to reduce FBT infection.

Insights on foodborne zoonotic trematodes in freshwater snails in North and Central Vietnam.

Foodborne zoonotic trematode (FZT) infections are common neglected tropical diseases in Southeast Asia. Their complicated life cycles involve freshwater snails as intermediate hosts. A cross-sectional study was conducted in Yen Bai and Thanh Hoa provinces in North and Central Vietnam, to investigate the diversity of cercariae of potential FZT and to construct the phylogenetic relationship of trematode cercariae based on the Internal Transcribed Spacer 2 (ITS2) region. Among 17 snail species collected from various habitats, 13 were infected by 10 cercarial groups among which parapleurolophocercous, pleurolophocercous, and echinostome cercariae were of zoonotic importance. The monophyletic tree separated cercarial sequences into different groups following the description of the cercariae families in which Haplorchidae, Opisthorchiidae, Echinochasmidae, and Echinostomatidae are important families of FZT. The overall prevalence was different among snail species and habitats and showed a seasonal trend. Parapleurolophocercous and echinostome cercariae emerged as the most common cercariae in snails in Yen Bai, while monostome, echinostome, and megalura cercariae were most common in Thanh Hoa. Using a molecular approach, we identified Parafossarulus striatulus as the first intermediate snail host of Clonorchis sinensis in Thac Ba Lake. Melanoides tuberculata and Bithynia fuchsiana were we identified preferred intermediate snail hosts of a diverse range of trematode species including intestinal flukes (i.e., Haplorchis pumilio and Echinochasmus japonicus) in Yen Bai and Thanh Hoa, respectively.

Bluetongue, Schmallenberg - what is next? Culicoides-borne viral diseases in the 21st Century.

In the past decade, two pathogens transmitted by Culicoides biting midges (Diptera: Ceratopogonidae), bluetongue virus and Schmallenberg virus, have caused serious economic losses to the European livestock industry, most notably affecting sheep and cattle. These outbreaks of arboviral disease have highlighted large knowledge gaps on the biology and ecology of indigenous Culicoides species. With these research gaps in mind, and as a means of assessing what potential disease outbreaks to expect in the future, an international workshop was held in May 2013 at Wageningen University, The Netherlands. It brought together research groups from Belgium, France, Germany, Spain, Switzerland, United Kingdom and The Netherlands, with diverse backgrounds in vector ecology, epidemiology, entomology, virology, animal health, modelling, and genetics. Here, we report on the key findings of this workshop.

Zoonoses in pet birds: review and perspectives.

Pet birds are a not-so-well known veterinarian's clientship fraction. Bought individually or in couples, as families often do (which is a lucrative business for pet shops or local breeders) or traded (sometimes illegally) for their very high genetic or exotic value, these birds, commonly canaries, parakeets or parrots, are regularly sold at high prices. These animals, however, are potential carriers and/or transmitters of zoonotic diseases. Some of them could have an important impact on human health, like chlamydophilosis, salmonellosis or even highly pathogenic avian influenza A H5N1. This review paper, although non exhaustive, aims at enlightening, by the description of several cases of bird-human transmission, the risks encountered by bird owners, including children. Public health consequences will be discussed and emphasis will be made on some vector-borne diseases, known to be emergent or which are underestimated, like those transmitted by the red mite Dermanyssus gallinae. Finally, biosecurity and hygiene, as well as prevention guidelines will be developed and perspectives proposed.

Identification of hard ticks (Acari: Ixodidae) and seroprevalence to Theileria parva in cattle raised in North Kivu Province, Democratic Republic of Congo.

This study aimed to identify tick species and to determine their relationship with the Theileria parva seroprevalence in cattle raised under an extensive farming system in North Kivu Province, Democratic Republic of Congo in two agro-ecological zones namely medium (1,000-1,850 m) and high (>1,850 m) altitude. Among the 3,215 ticks collected on 482 animals, from February to April 2009, Rhipicephalus appendiculatus (64.26 %), the main vector of T. parva, was the most abundant species followed by Rhipicephalus decoloratus (35.49 %) and Amblyomma variegatum (0.25 %). The mean burden of R. appendiculatus tick per infested animal appeared significantly higher at medium (6.5 ± 0.22 ticks) than at high (0.07 ± 0.3 ticks) altitude (P < 0.05). However, an indirect fluorescent antibody test carried out on 450 blood samples revealed a global T. parva seroprevalence of 43 % (95 % CI: 38-47) which was not significantly (P > 0.05) different between medium (48.4 %; 95 % CI: 38-49) and high (41.9 %; 95 % CI: 35-49) altitude. These relatively low seroprevalences suggest that there is a state of endemicity to T. parva infection in the study area. The presence of the tick vector on animals was associated with an increased risk of being seropositive to T. parva infection (odds ratio = 2.04; 95 % CI: 1.8-2.3; P < 0.001). The results suggest the need for a longitudinal study to investigate the seasonal dynamics of tick species and T. parva infection. The rate of tick infection should also be evaluated in order to determine the intensity of T. parva transmission to cattle.

Morphological and molecular characterization of lymnaeid snails and their potential role in transmission of Fasciola spp. in Vietnam.

Freshwater snails of the family Lymnaeidae play an important role in the transmission of fascioliasis worldwide. In Vietnam, 2 common lymnaeid species, Lymnaea swinhoei and Lymnaea viridis, can be recognized on the basis of morphology, and a third species, Lymnaea sp., is known to exist. Recent studies have raised controversy about their role in transmission of Fasciola spp. because of confusion in identification of the snail hosts. The aim of this study is, therefore, to clarify the identities of lymnaeid snails in Vietnam by a combination of morphological and molecular approaches. The molecular analyses using the second internal transcribed spacer (ITS2) of the nuclear ribosomal DNA clearly showed that lymnaeids in Vietnam include 3 species, Austropeplea viridis (morphologically identified as L. viridis), Radix auricularia (morphologically identified as L. swinhoei) and Radix rubiginosa (morphologically identified as Lymnaea sp.). R. rubiginosa is a new record for Vietnam. Among them, only A. viridis was found to be infected with Fasciola spp. These results provide a new insight into lymnaeid snails in Vietnam. Identification of lymnaeid snails in Vietnam and their role in the liver fluke transmission should be further investigated.

Longitudinal field study on bovine Babesia spp. and Anaplasma phagocytophilum infections during a grazing season in Belgium.

Anaplasmosis and babesiosis are major tick-borne diseases with a high economic impact but are also a public health concern. Blood samples collected in the spring, summer, and autumn of 2010 from 65 cows in seven different farms in Belgium were monitored with an indirect immunofluorescence antibody test to assess seroprevalence against these pathogens. Seroprevalences to Babesia spp. were measured as 10.7%, 20%, and 12.3% in spring, summer, and autumn, respectively, whereas seroprevalences to Anaplasma phagocytophilum were 30.8%, 77%, and 56.9%, respectively. A total of 805 Ixodes ricinus ticks were collected at the same time from both cattle (feeding ticks) and grazed pastures (questing ticks). The infection level of ticks, assessed by PCR assay, for Babesia spp. DNA was 14.6% and 7.9% in feeding and questing ticks, respectively, whereas 21.7% and 3% of feeding and questing ticks were found be positive for A. phagocytophilum cDNA. Fifty-five PCR-positive samples were identified by sequencing as Babesia sp. EU1, of which five from feeding ticks were positive for both A. phagocytophilum and Babesia sp. EU1. The high density of wild cervids in the study area could explain these observations, as deer are considered to be the main hosts for adults of I. ricinus. However, the absence of Babesia divergens both in feeding and questing ticks is surprising, as the study area is known to be endemic for cattle babesiosis. Increasing cervid populations and comorbidity could play an import role in the epidemiology of these tick-borne diseases.

Foodborne zoonotic trematode infections in Yen Bai, Vietnam: a situational analysis on knowledge, attitude, and practice (KAP) and risk behaviors.

Introduction: Foodborne Zoonotic Trematode Infections (FZTi) are neglected tropical diseases of public health concern in Vietnam. The transmission of FZTi is linked to human behavior patterns. The aims to investigate the knowledge, attitude, and practices regarding FZTi among local people. Methods: A cross-sectional study was conducted using a mixed method, which included a baseline survey and in-depth interviews. 375 participants were interviewed face-to-face in the survey, and 27 participants had the in-depth interviews. Results: The results showed that 36.3% passed the knowledge assessment, 86.7% passed the attitude assessment, and 24% passed the practical assessment. There were differences in average knowledge scores among gender (men higher than women, p = 0.006), ethnicities (Kinh higher than Dao and Tay, p < 0.001), and educational level (higher education, higher knowledge score, p < 0.001). There were differences in the frequency of eating raw fish between men and women (men higher than women, p < 0.001), and in the average practice score between men and women (women higher than men, p = 0.028). Eating raw fish and/or undercooked fish, raw vegetables, and drinking untreated water from Thac Ba lake in Yen Bai province were identified as FZTi risk behaviors. The occurrence of these risky habits can be explained by the lack of knowledge on FZTi, poor economic conditions and typical social features of local people. Conclusion: The current knowledge of local people in regard to safe eating practices is poor. They keep performing unsafe practices, which lead to infection with FZTi; therefore, an integrated control of FZTi is essential.

Effect of a commercial paratyphus vaccine on the development of pigeon circovirus infection in young pigeons (Columba livia domestica).

Infection with pigeon circovirus (PiCV) has been associated with young pigeon disease syndrome (YPDS), which is considered to be a multifactorial disease. The factors that determine whether birds succumb to clinical disease are not known. To evaluate the potential effect of vaccination with a commercial paratyphus vaccine on the progression of PiCV infection in young pigeons, forty 6-week-old pigeons naturally infected with PiCV were randomly assigned to two equal groups. The pigeons of one group were vaccinated at 6 and 9 weeks of age, and pigeons of the second group were unvaccinated controls. Cloacal swab and blood samples collected from all the birds were tested for PiCV by polymerase chain reaction (PCR) analysis. Three weeks after the second vaccination, all pigeons were euthanatized, and tissues were collected for PiCV PCR analysis and histopathologic evaluation. No significant difference in the number of PCR-positive cloacal swab and blood samples was found between the vaccinated and control pigeons. Positive PCR results in tissue samples also were not significantly different between the groups, with 18 positive samples in vaccinated birds (90%) and 16 in control birds (80%). Characteristic botryoid inclusions were detected in more vaccinated than control pigeons, but this difference was not significant. In this study, vaccination with a commercial paratyphus vaccine was not a risk factor for development of young pigeon disease syndrome.

Prevalence and risk factors associated with Clonorchis sinensis infections in rural communities in northern Vietnam.

BACKGROUND: Clonorchiasis, caused by the fish-borne trematode Clonorchis sinensis, is a neglected tropical disease and a public health issue in endemic countries. In Vietnam, an in-depth analysis of risk factors for the condition is missing up to now. This study aimed to determine the prevalence of C. sinensis infection and associated risk factors in rural communities in northern Vietnam. METHODOLOGY/PRINCIPAL FINDINGS: A cross-sectional survey was conducted in 4 communes in Yen Bai and Thanh Hoa provinces where clonorchiasis is known to be present and raw fish consumption is a common. Using a simple random sampling approach, stool was collected from 841 participants over 6 years old for coprological examination, and a questionnaire measured knowledge, attitudes, and practices with regard to clonorchiasis in 757 participants over 15 years old. Univariable and multivariable logistic regression models were run to identify risk factors for infection with C. sinensis. The overall prevalence of C. sinensis infection was 40.4%, with commune prevalences ranging between 26.5% and 53.3%. In the final model, males were significantly more likely to be infected with C. sinensis (OR 2.00; 95% CI 1.31-3.05). Recent (i.e. last year) consumption of raw fish (OR 8.00, 95% CI 4.78-13.36), low education level (OR 5.57; 95% CI 2.37-13.07), lack of treatment (OR 1.82, 95% CI 1.15-2.89), being between 19 to 39 years old (OR 6.46; 95% CI 1.25-33.37), and the presence of an unhygienic toilet (OR 2.74, 95% CI 1.53-4.92) were significantly associated with C. sinensis infection. CONCLUSION/SIGNIFICANCE: This study demonstrated a high prevalence of C. sinensis infection in rural communities in northern Vietnam. Thus, control measures including, mass drug administration for those communes should be applied to reduce the prevalence. Moreover, specific health education activities should be developed for risk groups in C. sinensis endemic areas.

Subversion of complement by hematophagous parasites.

The complement system is a crucial part of innate and adaptive immunity which exerts a significant evolutionary pressure on pathogens. It has selected for those pathogens, mainly microorganisms but also parasites, that have evolved countermeasures. The characterization of how pathogens evade complement attack is a rapidly developing field of current research. In recent years, multiple complement evasion strategies have been characterized. In this review, we focus on complement escape mechanisms expressed by hematophagous parasites, a heterogeneous group of metazoan parasites that share the property of ingesting the whole blood of their host. Complement inhibition is crucial for parasite survival within the host tissue or to facilitate blood feeding. Finally, complement inhibition by hematophagous parasites may also contribute to their success as pathogen vectors.

Evaluation of a SRS2 sandwich commercial enzyme-linked immunosorbent assay for the detection of anti-Neospora caninum antibodies in bovine and canine sera.

Neospora caninum is a parasite responsible for abortion in cows and neuromuscular disease in dogs. Serology is the most widely used technique to evaluate the prevalence of N. caninum in different host populations. A sandwich enzyme-linked immunosorbent assay (ELISA) based on the use of an anti-SRS2 monoclonal antibody was evaluated against the indirect fluorescent antibody test for 100 canine sera and against a well-characterized ELISA for 102 bovine sera. In cattle sera, the relative sensitivity and relative specificity were 100%. In dog sera, the relative specificity and relative sensitivity were 94% and 86%, respectively. The kappa value was 1 for bovine sera and 0.77 for canine sera. The seroprevalence was 3.9% in bovine sera and 21-23% in canine sera. The SRS2 sandwich ELISA was considered a valuable tool in both species.

Secreted subtilisins of Microsporum canis are involved in adherence of arthroconidia to feline corneocytes.

Microsporum canis is a pathogenic fungus that causes a superficial cutaneous infection called dermatophytosis, mainly in cats and humans. The mechanisms involved in adherence of M. canis to epidermis have never been investigated. Here, a model was developed to study the adherence of M. canis to feline corneocytes through the use of a reconstructed interfollicular feline epidermis (RFE). In this model, adherence of arthroconidia to RFE was found to be time-dependent, starting at 2 h post-inoculation and still increasing at 6 h. Chymostatin, a serine protease inhibitor, inhibited M. canis adherence to RFE by 53%. Moreover, two mAbs against the keratinolytic protease subtilisin 3 (Sub3) inhibited M. canis adherence to RFE by 23%, suggesting that subtilisins, and Sub3 in particular, are involved in the adherence process.

Secreted dipeptidyl peptidases as potential virulence factors for Microsporum canis.

Dermatophytoses caused by Microsporum canis are frequently encountered in cats and dogs; they are highly contagious and readily transmissible to humans. In this study, two single genes, respectively coding for dipeptidyl peptidases IV and V (DppIV and DppV), were isolated and characterized. Both proteins share homology with serine proteases of the S9 family, some of which display properties compatible with implication in pathogenic processes. Both genes are expressed in vivo in experimentally infected guinea-pigs and in naturally infected cats, and when the fungus is grown on extracellular matrix proteins as the sole nitrogen and carbon source. DppIV and V were produced as active recombinant proteases in the yeast Pichia pastoris; the apparent molecular weight of rDppV is 83 kDa, whereas rDppIV appears as a doublet of 95 and 98 kDa. Like other members of its enzymatic subfamily, rDppIV has an unusual ability to cleave Pro-X bonds. This activity does not enhance the solubilization of keratin by fungal secreted endoproteases, and the protease probably acts solely on small soluble peptides. RDppV showed no ability to induce delayed-type hypersensitivity (DTH) skin reactions in guinea-pigs, despite the known immunogenic properties of homologous proteins.

Detection of Neospora caninum in dog organs using real time PCR systems.

Neospora caninum is a parasite responsible for paresis in dogs. The dog can harbour enkysted parasites in several organs. The detection of N. caninum was performed using 3 different real time PCR systems all amplifying the NC5 DNA region. One system was based on Sybrgreen, one on Plexor technology and the last on Taqman probe. Comparison of the three methods indicated that the detection limit was 1 equivalent genome on pure DNA but that this detection limit increased in the presence of foreign DNA using the Sybrgreen and Plexor systems. Therefore, the Taqman system was chosen to detect N. caninum in liver and spleen of naturally infected dogs. The overall prevalence was 32.2%. Comparison between PCR results and serological results using IFAT showed that among the 28 PCR positive dogs only 9 were seropositive and that 8 seropositive dogs were PCR negative. Therefore serology can underestimate the real carriage in dogs. However, PCR methods must be improved in terms of sensitivity and inhibition problems.

Reconstructed interfollicular feline epidermis as a model for the screening of antifungal drugs against Microsporum canis.

A fully differentiated reconstructed interfollicular feline epidermis (RFE) was recently developed in vitro. It was shown to be relevant for the study of Microsporum canis-epidermal interactions. In this study, RFE was evaluated as a potential model for the in vitro screening of drugs against M. canis. As a preliminary step, the minimum inhibitory concentration of miconazole nitrate against M. canis IHEM 21239 grown on Sabouraud's dextrose agar was determined to be 0.3 microg mL(-1). RFE grown at the air-liquid interface was cultured for 24 h in RFE culture medium, supplemented with either miconazole (range 0.1-1 microg mL(-1)) or its solvent (dimethylsulfoxide). Then, RFE was inoculated in triplicate with 1 x 10(5 )M. canis arthroconidia and incubated for five additional days. To evaluate fungal growth, RFE was processed for routine histopathology, three serial sections being performed across the block at 100 microm intervals. No fungal growth was detected invading or on the surface of infected RFE in the presence of miconazole concentrations equal to or higher than 0.3 microg mL (final concentration in the culture medium). This study demonstrates that RFE is an adequate model for the in vitro screening of drugs against M. canis and potentially against other skin pathogens.