Next Article in Journal
First Report of Enterocytozoon hepatopenaei Infection in Giant Freshwater Prawn (Macrobrachium rosenbergii de Man) Cultured in the Republic of Korea
Next Article in Special Issue
Brazilian Horses from Bahia State Are Highly Infected with Sarcocystis bertrami
Previous Article in Journal
Gene and Protein Accumulation Changes Evoked in Porcine Aorta in Response to Feeding with Two Various Fructan Sources
Previous Article in Special Issue
Neospora spp. Seroprevalence and Risk Factors for Seropositivity in Apparently Healthy Horses and Pregnant Mares
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Animals
Volume 12
Issue 22
10.3390/ani12223148
animals-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Molecular Detection and Multilocus Genotyping of Giardia duodenalis in Pigs in Fujian Province, Southeastern China

by
Fu-Rong Zhao
1,2,3,
Ning Zhang
2,
Wen-Yuan Miao
2,
Ran Wu
2,
Lin-Lin Cui
2,
Cui-Qin Huang
3,4,* and
Dong-Hui Zhou
2,*
1
Fujian Key Laboratory on Conservation and Sustainable Utilization of Marine Biodiversity, Fuzhou Institute of Oceanography, Minjiang University, Fuzhou 350108, China
2
Key Laboratory of Fujian-Taiwan Animal Pathogen Biology, College of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, China
3
Fujian Provincial Key Laboratory for the Prevention & Control of Animal Infectious Diseases & Biotechnology, Longyan University, Longyan 364012, China
4
Engineering Research Center for the Prevention and Control of Animal Original Zoonosis, Fujian Province University & College of Life Science, Longyan 364012, China
*
Authors to whom correspondence should be addressed.
Animals 2022, 12(22), 3148; https://doi.org/10.3390/ani12223148
Submission received: 22 September 2022 / Revised: 1 November 2022 / Accepted: 2 November 2022 / Published: 14 November 2022
(This article belongs to the Special Issue Parasite Epidemiology and Molecules Identification in Wild and Domestic Animals)
Browse Figures
Versions Notes

Abstract

:

Simple Summary

Giardia duodenalis is a world-widely distribution intestinal protozoan parasite that can infect human and a broad range of mammals. It can cause a number of clinical symptoms including diarrhoea, abdominal pain, vomiting, severe dehydration and anemia etc. in animals and humans. Eight genetic assemblages (A to H) have been identified in G. duodenalis based on molecular analysis. In this study, the infection rates and genotypes of G. duodenalis in swine are investigated in Fujian province, southeastern China. G. duodenalis assemblages E was found in pigs in the present study. These results provide epidemiological data for giardiasis prevention, control and public health.

Abstract

Giardia duodenalis, an intestinal parasite, is widely distributed in humans and various animals, such as pigs, cattle and cats. The clinical symptoms of giardiasis are characterized as including abdominal pain, acute or chronic diarrhea, and bloating and weight loss in humans and animals, leading to public and veterinary health problems worldwide. However, the prevalence and genotypes of G. duodenalis in pigs in Fujian Province, southeastern China, have not been reported. In the present study, 725 fecal samples were collected from six cities (Fuqing, Putian, Nanping, Longyan, Sanming, Zhangzhou) in Fujian Province and analyzed for G. duodenalis prevalence and genotypes using nested PCR targeting the beta-giardin (bg), glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi) genes. The results shown that total occurrence rate of G. duodenalis was 26.9% (195/725) in pigs, with significant differences in the prevalence among different regions (χ2 = 86.508, p < 0.05) and groups (χ2 = 12.748, p < 0.05). 195, 11 and 6 samples were detected at the bg, tpi and gdh loci, respectively. Each one belonged to a subtype of assemblage E and was analyzed using sequences obtained in this study. Based on phylogenetic analyses of sequences from the three genetic loci, only one MLG E1 was found. The results indicated that pigs may present a potential zoonotic risk of spreading G. duodenalis infection from animals to humans in this area. The findings of the present study also provide basic data for the prevention and control of G. duodenalis infection in pigs and humans in China.

1. Introduction

Giardia duodenalis (syn, G. intestinalis or G. lambila) is an intestinal protozoan parasite distributed worldwide and transmitted through the fecal-oral route [1]. Humans and animals are infected by ingesting G. duodenalis cyst-contaminated water and food or by direct touch with infected animals, which causes abdominal pain, acute or chronic diarrhea, and bloating and weight loss [2,3]. Acute giardiasis develops after an incubation period of 1 to 14 days (average of 7 days) and usually lasts 1 to 3 weeks. Repeated samplings are usually required on different days during diagnosis because Giardia cysts and trophozoites are not always present in the feces. So far, no effective vaccines have been able to prevent and control giardiasis. There are some antimicrobial drugs for treatment of giardiasis, such as metronidazole, nitazoxanide, paromomycin, tinidazole, quinacrine and furazolidone. However, these drugs can cause side effects (e.g., metallic taste) and drug resistance in many poor patients [1,4]. Some studies have reported a high prevalence and disease burden of G. duodenalis infection in animals, such as pigs, cattle, dogs, cats and wildlife [5,6,7]. Thus, G. duodenalis has a significant health impact on both humans and animals.
To date, eight assemblages (A–H) have been identified in G. duodenalis based on genetic analysis. Among them, assemblages A and B are found in both humans and a variety of mammals, and the remaining six assemblages are strongly host-specific [8,9]. It is commonly believed that humans are infected only with assemblages A and B, but the sporadic infection by assemblages C, D, E and F have also been isolated from humans [10,11,12,13]. At present, there are prevalence reports and genotype analyses of G. duodenalis infection in pigs all over the world. In the published studies, assemblages A, B and E were identified in pigs, with assemblage E being the predominant genotype [14,15,16,17].
Due to the inadequate genetic characterization of G. duodenalis using only a single gene, and even sometimes a limited sensitivity, genotyping of G. duodenalis is most commonly achieved by the amplification of several target genes, such as the small subunit of ribosomal RNA (SSU rRNA), beta-giardin (bg), triose phosphate isomerase (tpi) and glutamate dehydrogenase (gdh) [1,8]. In recent years, multilocus genotyping (MLG) based on the three genes (bg, tpi and gdh) was developed and has been applied to provide more genetic information and contribute to understanding possible zoonotic transmission linkages [18,19]. However, limited data on G. duodenalis infection in pigs in China is available. Thus, the current study was intended to characterize G. duodenalis in pigs in Fujian Province and to assess the public health potential of G. duodenalis in pigs.

2. Materials and Methods

2.1. Specimen Collection

In total, 725 fresh fecal samples were collected from six regions (121 from Fuqing, 146 from Putian, 139 from Nanping, 120 from Longyan, 118 from Sanming and 81 from Zhangzhou) in Fujian Province (Location: 24′ N to 28′ N and 116′ E to 121′ E), southeastern China (Figure 1). These samples were collected from industry farms with more than 500 breeding sows. The distribution of samples by type was as follows: 111 samples from suckling pigs, 127 samples from weaned pigs, 104 samples from nursery pigs, 298 samples from sows, 28 samples from boars and 57 samples from fattening pigs. All the fecal samples were placed in clean plastic bags and preserved in 2.5% potassium dichromate (4 °C) prior to DNA extraction.

2.2. DNA Extraction and Nested PCR Amplification

Approximately 200 mg of feces per sample was placed in a sterile centrifuge tube, and the potassium dichromate solution was washed three times with distilled water. Genomic DNA was extracted using the commercial E.Z.N.A® Stool DNA kit (Omega Biotek Inc., Norcross, GA, USA) according to the manufacturer’s protocol. G. duodenalis in fecal specimens was determined using nested PCR analysis of the beta-giardin (bg) gene. The primers and amplification used in this study have been previously described [1,7]. The final amplification products were subjected to electrophoresis in 1% agarose gel stained with Gold ViewTM (Solar bio Co., Ltd., Beijing, China) and were visualized under UV light.

2.3. Sequence Analysis

G. duodenalis positive secondary PCR products from the bg, tpi and gdh genes were sequenced bidirectionally with secondary PCR primers using an ABI PRISMTM 3730 XL DNA Analyzer (Applied Biosystems, Foster City, CA, USA) at the Sune Biotech Co., Ltd. (Fuzhou, China). Then, the obtained DNA sequences were aligned with references downloaded from the National Center for Biotechnology Information (NCBI) website (http://www.ncbi.nlm.nih.gov/BLAST (accessed on 16 August 2022)) using the software Clustal X v.2.1 (http://www.clustal.org/ (accessed on 16 August 2022)) to identify G. duodenalis assemblages/subtypes.

2.4. Phylogenetic Analysis

MEGA X version 10.1.7 (http://www.megasoftware.net (accessed on 16 August 2022)) software was used to perform phylogenetic analyses using the neighbor-joining method with the Kimura 2-parameter model. Bootstrap analyses with 1000 replicates were used to assess the robustness of cluster formation.

2.5. Statistical Analysis

Differences in infection rate between husbandry parameters, including groups and locations, were compared with the chi-square test (χ2) using the software SPSS 22.0 (IBM Corp., New York, NY, USA). Values were considered to be statistically significant when p < 0.05.

3. Results

From a total of 725 fecal specimens in this study, 195 (26.9%) fecal samples tested positive for G. duodenalis based on the beta-giardin (bg) gene. The highest infection rate was seen in specimens from Sanming (49.2%, 58/118), followed by Nanping (36.7%, 51/139), Fuqing (36.4%, 44/121), Zhangzhou (22.2%, 18/81), Longyan (10.8%, 13/120) and Putian (7.5%, 11/146), and was statistically significant (χ2 = 86.508, p < 0.05). Among the six groups of pigs, suckling pigs (35.1%, 39/111) had the highest infection rate, followed by weaned pigs (29.1%, 37/127), boars (28.8%, 8/28), sows (27.2%, 81/298), nursery pigs (23.1%, 24/104) and fattening pigs (10.5%, 6/57), and the difference was statistically significant (χ2 = 12.748, p < 0.05) (Table 1).
Based on the bg gene locus, all 195 G. duodenalis positive samples belonged to assemblage E, among which subtypes E5 and E15 were found, and also showed 100% nucleotide identity with a cattle-derived sequence (GenBank accession number: MH158458) and a Tibetan sheep-derived sequence (GenBank accession number: KY633473) respectively. In total, 11 and 6 Giardia-positive samples belonging to assemblage E15 were identified at the tpi and gdh gene loci, respectively, and showed 100% nucleotide identity with a pig-derived sequence (GenBank accession number: KJ668136) and another pig-derived sequence (GenBank accession number: KJ668145), respectively. Unfortunately, attempts to access data at the tpi and gdh gene loci failed in all assemblage E5 samples. Phylogenetic analyses show that all sequences generated in this study at the gdh, bg and tpi loci were grouped in well-defined clusters with suitable references and matched previously published sequences retrieved from GenBank (Figure 2, Figure 3 and Figure 4).
Six samples were amplified at three loci at the same time and only one MLG E1 was formed (Table 2).

4. Discussion

G. duodenalis infection in pigs has been reported worldwide. In this study, the total infection rate of G. duodenalis was 26.9% (195/725), which was close to the infection rate detected using PCR based on the bg, tpi and gdh loci and using ELISA in Shanghai (26.88%) [20] and Nigeria (25.4%) [21] in 2019. The infection rate in the present study detected using PCR based on the SSU rRNA and bg loci was lower than the following that were reported: 31.1% in Australia in 2009 [22], 66.4% in Canada in 2011 [23] and 57.1% in the United Kingdom in 2014 [24]. However, the infection rate in the present study detected using microscopy and PCR based on the SSU rRNA, bg, tpi and gdh loci was higher than most reports on G. duodenalis: 17.4% in Denmark in 2006 [25], 7.4% in the United States in 1994 [26], 9.5% in Poland in 2015 [27], 8% in Shaanxi in 2017 [15], 14.8% in Korea in 2020 [28], 4.26% in Taiwan in 2021 [29] and 0.97% in Hubei in 2022 [30]. Many factors, such as testing methods, sample size, sampling time and geographic location, may provide reasons for the differences in G. duodenalis prevalence.
Infection rates were statistically significant among different groups (χ2 = 86.508, p < 0.05), with the highest prevalence in suckling pigs and the lowest in fattening pigs, which were similar to those of Australia and Denmark [22,31]. However, the study on the infection of G. duodenalis in Shaanxi Province showed that the infection rate of sows was the highest (10.5%) [15], which was consistent with the result in Lusaka, Zambia (53.3%) [32]. Multiple factors, such as loci amplified, climate, age structure and sample size, may lead to various infection rates.
The alignment of nucleotide sequences obtained in this investigation showed only one genotype of assemblage E in Fujian Province. The previous report shows that ungulates, including swine, are easily infected by assemblage E, which can also infect humans [10]. This indicates that assemblage E presents a risk for humans and animals. Study results about G. duodenalis in Shaanxi Province found that the main zoonotic genotype was assemblage A (20%) [15]. Assemblage E and A mixed infections were observed in Denmark [25]. The distribution of assemblages in different regions is slightly different, so further studies are needed on the epidemiology and genotypic transmission mechanism of human and swine G. duodenalis.
In total, six samples were amplified at three loci at the same time. Only one MLG E1 was formed. In Shaanxi Province, eight samples were amplified to form four MLGs at the three loci of bg, tpi and gdh. In addition, coinfection with assemblage A and assemblage E was identified in one fattening pig [15]. The results showed that polymorphism in G. duodenalis was different among regions. However, more data are needed on the specific mechanism due to few reports on the MLG typing of G. duodenalis in pigs.
G. duodenalis induces a strongly adaptive immune response in both humans and animals. After infection, a large number of parasitic-specific IgA are produced, and the CD4+ T cell response is helpful for the production of IgA and the control of infection [33]. Piglets do not become fully immune until they are about 4 weeks old after birth, which means they are less able to respond to the source of infection during the first month [34]. This is consistent with the highest rate of infection in suckling pigs in our survey. Despite the global spread of G. duodenalis infection, there is, unfortunately, currently no protective vaccine available for human use, and drug treatment regimens have had varying effects [35]. As an omnivorous and monogastric species, pigs are very similar to humans in terms of anatomical structure and immune system function. Studies have reported that the similarity between the immune systems of pigs and humans is more than 80% [36]. Therefore, pigs have many advantages as animal models of human diseases.

5. Conclusions

In conclusion, the total infection rate of G. duodenalis was 26.9% (195/725) in pigs in Fujian Province, southeastern China, and only one MLG was formed. To the best of our knowledge, this is the first report of G. duodenalis infection in pigs in Fujian Province. Considering that pigs are necessary economic animals in China, a better understanding of G. duodenalis in pigs will help to develop more targeted prevention and control measures, which have essential implications for the control of G. duodenalis infection in pigs and humans.

Author Contributions

D.-H.Z. and C.-Q.H. conceived and designed this study. F.-R.Z. and N.Z. performed the experiments. F.-R.Z. and W.-Y.M. drafted the manuscript. L.-L.C. and R.W. contributed reagents/materials/analysis tools. D.-H.Z. critically revised the manuscript. All authors have read and agreed to the published version of the manuscript.

Funding

This work supported by the Natural Science Foundation of Fujian Province of China (2021J011044), the Fujian Provincial Key Laboratory for Prevention and Control of Animal Infectious Diseases and Biotechnology (ZDSYS2020002) and the Open Fund of the Key Laboratory of Fujian Province Livestock Epidemic Prevention, Control and Biological Technology (2018KF02).

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Not applicable.

Conflicts of Interest

All authors declare that they have no conflict of interest.

References

  1. Feng, Y.; Xiao, L. Zoonotic potential and molecular epidemiology of Giardia species and giardiasis. Clin. Microbiol. Rev. 2011, 24, 110–140. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  2. Liu, L.; Wei, Z.; Fang, R.; Li, X.; Li, W. Giardia duodenalis induces extrinsic pathway of apoptosis in intestinal epithelial cells through activation of TNFR1 and K63 de-ubiquitination of RIP1 in vitro. Microb. Pathog. 2020, 149, 104315. [Google Scholar] [CrossRef] [PubMed]
  3. Certad, G.; Viscogliosi, E.; Chabé, M.; Cacciò, S.M. Pathogenic Mechanisms of Cryptosporidium and Giardia. Trends Parasitol. 2017, 33, 561–576. [Google Scholar] [CrossRef] [PubMed]
  4. Vivancos, V.; González-Alvarez, I.; Bermejo, M. Giardiasis: Characteristics, Pathogenesis and New Insights about Treatment. Curr. Top. Med. Chem. 2018, 18, 1287–1303. [Google Scholar] [CrossRef] [PubMed]
  5. Thompson, R.C. The zoonotic significance and molecular epidemiology of Giardia and giardiasis. Vet. Parasitol. 2004, 126, 15–35. [Google Scholar] [CrossRef]
  6. Appelbee, A.J.; Thompson, R.C.; Olson, M.E. Giardia and Cryptosporidium in mammalian wildlife-current status and future needs. Trends Parasitol. 2005, 21, 370–376. [Google Scholar] [CrossRef]
  7. Geurden, T.; Vandenhoute, E.; Pohle, H.; Casaert, S.; De Wilde, N.; Vercruysse, J.; Claerebout, E. The effect of a fenbendazole treatment on cyst excretion and weight gain in calves experimentally infected with Giardia duodenalis. Vet. Parasitol. 2010, 169, 18–23. [Google Scholar] [CrossRef]
  8. Lalle, M.; Pozio, E.; Capelli, G.; Bruschi, F.; Crotti, D.; Cacciò, S.M. Genetic heterogeneity at the beta-giardin locus among human and animal isolates of Giardia duodenalis and identification of potentially zoonotic subgenotypes. Int. J. Parasitol. 2005, 35, 207–213. [Google Scholar] [CrossRef]
  9. Einarsson, E.; Ma’ayeh, S.; Svärd, S.G. An up-date on Giardia and giardiasis. Curr. Opin. Microbiol. 2016, 34, 47–52. [Google Scholar] [CrossRef]
  10. Fantinatti, M.; Bello, A.R.; Fernandes, O.; Da-Cruz, A.M. Identification of Giardia lamblia Assemblage E in Humans Points to a New Anthropozoonotic Cycle. J. Infect. Dis. 2016, 214, 1256–1259. [Google Scholar] [CrossRef]
  11. Foronda, P.; Bargues, M.D.; Abreu-Acosta, N.; Periago, M.V.; Valero, M.A.; Valladares, B.; Mas-Coma, S. Identification of genotypes of Giardia intestinalis of human isolates in Egypt. Parasitol. Res. 2008, 103, 1177–1181. [Google Scholar] [CrossRef] [PubMed]
  12. Minetti, C.; Lamden, K.; Durband, C.; Cheesbrough, J.; Fox, A.; Wastling, J.M. Determination of Giardia duodenalis assemblages and multi-locus genotypes in patients with sporadic giardiasis from England. Parasites Vectors 2015, 8, 444. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  13. Zahedi, A.; Field, D.; Ryan, U. Molecular typing of Giardia duodenalis in humans in Queensland—First report of Assemblage E. Parasitology 2017, 144, 1154–1161. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  14. Li, W.; Deng, L.; Wu, K.; Huang, X.; Song, Y.; Su, H.; Hu, Y.; Fu, H.; Zhong, Z.; Peng, G. Presence of zoonotic Cryptosporidium scrofarum, Giardia duodenalis assemblage A and Enterocytozoon bieneusi genotypes in captive Eurasian wild boars (Sus scrofa) in China: Potential for zoonotic transmission. Parasites Vectors 2017, 10, 10. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  15. Wang, S.S.; Yuan, Y.J.; Yin, Y.L.; Hu, R.-S.; Song, J.-K.; Zhao, G.-H. Prevalence and multilocus genotyping of Giardia duodenalis in pigs of Shaanxi Province, northwestern China. Parasites Vectors 2017, 10, 490. [Google Scholar] [CrossRef] [Green Version]
  16. Wang, H.; Zhang, Y.; Wu, Y.; Li, J.; Qi, M.; Li, T.; Wang, J.; Wang, R.; Zhang, S.; Jian, F.; et al. Occurrence, Molecular Characterization, and Assessment of Zoonotic Risk of Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi in Pigs in Henan, Central China. J. Eukaryot Microbiol. 2018, 65, 893–901. [Google Scholar] [CrossRef] [PubMed]
  17. Jing, B.; Zhang, Y.; Xu, C.; Li, D.; Xing, J.; Tao, D.; Zhang, L.; Qi, M.; Wang, H. Detection and genetic characterization of Giardia duodenalis in pigs from large-scale farms in Xinjiang, China. Parasite 2019, 26, 53. [Google Scholar] [CrossRef] [Green Version]
  18. Cacciò, S.M.; Beck, R.; Lalle, M.; Marinculic, A.; Pozio, E. Multilocus genotyping of Giardia duodenalis reveals striking differences between assemblages A and B. Int. J. Parasitol. 2008, 38, 1523–1531. [Google Scholar] [CrossRef]
  19. Scorza, A.V.; Ballweber, L.R.; Tangtrongsup, S.; Panuska, C.; Lappin, M.R. Comparisons of mammalian Giardia duodenalis assemblages based on the β-giardin, glutamate dehydrogenase and triose phosphate isomerase genes. Vet. Parasitol. 2012, 189, 182–188. [Google Scholar] [CrossRef]
  20. Liu, H.; Xu, N.; Yin, J.; Yuan, Z.; Shen, Y.; Cao, J. Prevalence and multilocus genotyping of potentially zoonotic Giardia duodenalis in pigs in Shanghai, China. Parasitology 2019, 146, 1199–1205. [Google Scholar] [CrossRef]
  21. Akinkuotu, O.A.; Takeet, M.I.; Otesile, E.B.; Olufemi, F.; Greenwood, S.J.; McClure, J.T. Prevalence and multilocus genotypes of Giardia duodenalis infecting pigs in Ogun state, Nigeria. Infect. Genet. Evol. 2019, 70, 53–60. [Google Scholar] [CrossRef] [PubMed]
  22. Armson, A.; Yang, R.; Thompson, J.; Johnson, J.; Reid, S.; Ryan, U.M. Giardia genotypes in pigs in Western Australia: Prevalence and association with diarrhea. Exp. Parasitol. 2009, 121, 381–383. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  23. Farzan, A.; Parrington, L.; Coklin, T.; Cook, A.; Pintar, K.; Pollari, F.; Friendship, R.; Farber, J.; Dixon, B. Detection and characterization of Giardia duodenalis and Cryptosporidium spp. on swine farms in Ontario, Canada. Foodborne Pathog. Dis. 2011, 8, 1207–1213. [Google Scholar] [CrossRef] [PubMed]
  24. Minetti, C.; Taweenan, W.; Hogg, R.; Featherstone, C.; Randle, N.; Latham, S.M.; Wastling, J.M. Occurrence and diversity of Giardia duodenalis assemblages in livestock in the UK. Transbound. Emerg. Dis. 2014, 61, e60–e67. [Google Scholar] [CrossRef] [Green Version]
  25. Maddox-Hyttel, C.; Langkjaer, R.B.; Enemark, H.L.; Vigre, H. Cryptosporidium and Giardia in different age groups of Danish cattle and pigs--occurrence and management associated risk factors. Vet. Parasitol. 2006, 141, 48–59. [Google Scholar] [CrossRef]
  26. Xiao, L.; Herd, R.P.; Bowma, L.G. Prevalence of Cryptosporidium and Giardia infections on two Ohio pig farms with different management systems. Vet. Parasitol. 1994, 52, 331–336. [Google Scholar] [CrossRef]
  27. Stojecki, K.; Sroka, J.; Cencek, T.; Dutkiewicz, J. Epidemiological survey in Łęczyńsko-Włodawskie Lake District of eastern Poland reveals new evidence of zoonotic potential of Giardia intestinalis. Ann. Agric. Environ. Med. 2015, 22, 594–598. [Google Scholar] [CrossRef]
  28. Lee, H.; Jung, B.; Lim, J.S.; Seo, M.G.; Lee, S.H.; Choi, K.H.; Hwang, M.H.; Kim, T.H.; Kwon, O.D.; Kwak, D. Multilocus genotyping of Giardia duodenalis from pigs in Korea. Parasitol. Int. 2020, 78, 102154. [Google Scholar] [CrossRef]
  29. Lam, H.Y.P.; Chen, T.T.; Tseng, Y.C.; Chang, K.-C.; Yang, T.-H.; Peng, S.-Y. Detection and genotyping of Giardia duodenalis from cattle and pigs in Hualien country, Eastern Taiwan. J. Microbiol. Immunol. Infect. 2021, 54, 718–727. [Google Scholar] [CrossRef]
  30. Li, D.; Deng, H.; Zheng, Y.; Zhang, H.; Wang, S.; He, L.; Zhao, J. First Characterization and Zoonotic Potential of Cryptosporidium spp. and Giardia duodenalis in Pigs in Hubei Province of China. Front. Cell. Infect. Microbiol. 2022, 12, 949773. [Google Scholar] [CrossRef]
  31. Petersen, H.H.; Jianmin, W.; Katakam, K.K.; Mejer, H.; Thamsborg, S.M.; Dalsgaard, A.; Olsen, A.; Enemark, H.L. Cryptosporidium and Giardia in Danish organic pig farms: Seasonal and age-related variation in prevalence, infection intensity and species/genotypes. Vet. Parasitol. 2015, 214, 29–39. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  32. Siwila, J.; Mwape, K.E. Prevalence of Cryptosporidium spp. and Giardia duodenalis in pigs in Lusaka, Zambia. Onderstepoort J. Vet. Res. 2012, 79, a404. [Google Scholar] [CrossRef] [PubMed]
  33. Singer, S.M.; Fink, M.Y.; Angelova, V.V. Recent insights into innate and adaptive immune responses to Giardia. Adv. Parasitol. 2019, 106, 171–208. [Google Scholar] [CrossRef] [PubMed]
  34. Poonsuk, K.; Zimmerman, J. Historical and contemporary aspects of maternal immunity in swine. Anim. Health Res. Rev. 2018, 19, 31–45. [Google Scholar] [CrossRef] [PubMed]
  35. Ansell, B.R.; McConville, M.J.; Ma’ayeh, S.Y.; Dagley, M.J.; Gasser, R.B.; Svärd, S.G.; Jex, A.R. Drug resistance in Giardia duodenalis. Biotechnol. Adv. 2015, 33, 888–901. [Google Scholar] [CrossRef]
  36. Pabst, R. The pig as a model for immunology research. Cell Tissue Res. 2020, 380, 287–304. [Google Scholar] [CrossRef]
Animals 12 03148 g001 550
Figure 1. Geographic distribution of sampling sites in Fujian Province, southeastern China.
Figure 1. Geographic distribution of sampling sites in Fujian Province, southeastern China.
Animals 12 03148 g001
Animals 12 03148 g002 550
Figure 2. Phylogenetic relationships of Giardia duodenalis isolates obtained using the neighbor-joining analysis of the beta-giardin (bg) nucleotide sequences. Bootstrap values >50% from 1000 replicates are shown as nodes. Giardia muris was used as outgroup. Filled black circles represent bg sequences generated in this study. The scale bar indicates 0.01 nucleotide substitution/site.
Figure 2. Phylogenetic relationships of Giardia duodenalis isolates obtained using the neighbor-joining analysis of the beta-giardin (bg) nucleotide sequences. Bootstrap values >50% from 1000 replicates are shown as nodes. Giardia muris was used as outgroup. Filled black circles represent bg sequences generated in this study. The scale bar indicates 0.01 nucleotide substitution/site.
Animals 12 03148 g002
Animals 12 03148 g003 550
Figure 3. Phylogenetic relationships of Giardia duodenalis isolates obtained using the neighbor-joining analysis of the glutamate dehydrogenase (gdh) nucleotide sequences. Bootstrap values >50% from 1000 replicates are shown as nodes. Giardia ardeae was used as outgroup. Filled black circles represent bg sequences generated in this study. The scale bar indicates 0.2 nucleotide substitution/site.
Figure 3. Phylogenetic relationships of Giardia duodenalis isolates obtained using the neighbor-joining analysis of the glutamate dehydrogenase (gdh) nucleotide sequences. Bootstrap values >50% from 1000 replicates are shown as nodes. Giardia ardeae was used as outgroup. Filled black circles represent bg sequences generated in this study. The scale bar indicates 0.2 nucleotide substitution/site.
Animals 12 03148 g003
Animals 12 03148 g004 550
Figure 4. Phylogenetic relationships of Giardia duodenalis isolates obtained using the neighbor-joining analysis of the triose phosphate isomerase (tpi) nucleotide sequences. Bootstrap values >50% from 1000 replicates are shown as nodes. Giardia ardeae is used as outgroup. Filled black circles represent bg sequences generated in this study. The scale bar indicates 0.05 nucleotide substitution/site.
Figure 4. Phylogenetic relationships of Giardia duodenalis isolates obtained using the neighbor-joining analysis of the triose phosphate isomerase (tpi) nucleotide sequences. Bootstrap values >50% from 1000 replicates are shown as nodes. Giardia ardeae is used as outgroup. Filled black circles represent bg sequences generated in this study. The scale bar indicates 0.05 nucleotide substitution/site.
Animals 12 03148 g004
Table
Table 1. Prevalence of G. duodenalis infection in pigs in Fujian Province.
Table 1. Prevalence of G. duodenalis infection in pigs in Fujian Province.
VariableCategoryNo. IsolatesNo. Positive (%, 95% CI)Assemblage (Number)p-Value
GroupSuckling pigs11139 (35.1%, 26.9–44.4%)E (39)<0.05
Weaned pigs12737 (29.1%, 21.9–37.5%)E (37)
Nursery pigs10424 (23.1%, 16.0–32.1%)E (24)
Fattening pigs576 (10.5%, 4.9–21.1%)E (6)
Sows29881 (27.2%, 22.4–32.5%)E (81)
Boars288 (28.6%, 15.3–47.1%)E (8)
LocationFuqing City12144 (36.4%, 28.3–45.2%)E (44)<0.05
Nanping City13951 (36.7%, 29.1–44.9%)E (51)
Longyan City12013 (10.8%, 6.4–17.6%)E (13)
Putian City14611 (7.5%, 4.2–12.9%)E (11)
Sanming City11858 (49.2%, 40.3–58.1%)E (58)
Zhangzhou City8118 (22.2%, 14.5–32.4%)E (18)
Total725195 (26.9%, 23.8–30.2%)E (195)
Table
Table 2. Multilocus characterization of G. duodenalis isolates based on bg, tpi and gdh genes.
Table 2. Multilocus characterization of G. duodenalis isolates based on bg, tpi and gdh genes.
IsolateGenotype or SubtypeMLG Type
bgtpigdh
NP-68E15E1E1MLG E1
NP-69E15E1E1MLG E1
NP-71E15E1E1MLG E1
NP-73E15E1E1MLG E1
NP-79E15E1E1MLG E1
NP-81E15E1--
NP-85E15E1--
NP-86E15E1E1MLG E1
NP-70E15E1--
NP-78E15E1--
PT-YF3E15E1--
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Share and Cite

MDPI and ACS Style

Zhao, F.-R.; Zhang, N.; Miao, W.-Y.; Wu, R.; Cui, L.-L.; Huang, C.-Q.; Zhou, D.-H. Molecular Detection and Multilocus Genotyping of Giardia duodenalis in Pigs in Fujian Province, Southeastern China. Animals 2022, 12, 3148. https://doi.org/10.3390/ani12223148

AMA Style

Zhao F-R, Zhang N, Miao W-Y, Wu R, Cui L-L, Huang C-Q, Zhou D-H. Molecular Detection and Multilocus Genotyping of Giardia duodenalis in Pigs in Fujian Province, Southeastern China. Animals. 2022; 12(22):3148. https://doi.org/10.3390/ani12223148

Chicago/Turabian Style

Zhao, Fu-Rong, Ning Zhang, Wen-Yuan Miao, Ran Wu, Lin-Lin Cui, Cui-Qin Huang, and Dong-Hui Zhou. 2022. "Molecular Detection and Multilocus Genotyping of Giardia duodenalis in Pigs in Fujian Province, Southeastern China" Animals 12, no. 22: 3148. https://doi.org/10.3390/ani12223148

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop