Animals

2024

Jump to: 2023, 2022, 2021

15 pages, 5289 KiB

Open AccessArticle

Dirofilaria sp. and Blood Meal Analysis in Mosquitoes Collected in Vojvodina and Mačva, and the First Report of Setaria tundra (Issaitshikoff & Rajewskaya, 1928) in Serbia

by Sara Šiljegović, Théo Mouillaud, Davy Jiolle, Dušan Petrić, Aleksandra Ignjatović-Ćupina, Ana Vasić, Christophe Paupy and Mihaela Kavran

Animals 2024, 14(9), 1255; https://doi.org/10.3390/ani14091255 - 23 Apr 2024

Viewed by 1218

Abstract

Dirofilaria immitis and D. repens are the two most widespread and important species of mosquito-borne nematodes, posing a significant threat to veterinary health and particularly affecting canines and felines. While D. immitis causes cardiopulmonary dirofilariasis, D. repens causes subcutaneous infections in dogs and [...] Read more.

Dirofilaria immitis and D. repens are the two most widespread and important species of mosquito-borne nematodes, posing a significant threat to veterinary health and particularly affecting canines and felines. While D. immitis causes cardiopulmonary dirofilariasis, D. repens causes subcutaneous infections in dogs and other carnivores. Despite the extensive knowledge on these parasites, little is known about their natural vectors in Serbia. The parasite Setaria tundra, known to infect deer, has not yet been detected in Serbia but has been documented in neighboring countries. Thus, the aim of this study was to (i) further map out Dirofilaria sp. hotspots in the Vojvodina Province and detect S. tundra for the first time, (ii) detect positive mosquito species that can provide insights into how the nematodes spread in Serbia, and (iii) analyze the blood-fed female mosquitoes of species found to be infected, in order to identify the potential source of parasite infection. A total of 2902 female mosquitoes were collected across 73 locations during 2021 and 2022. Molecular biology methods, based on conventional PCR, were used to analyze non-blood-fed (2521 specimens) and blood-fed (381 specimens) female mosquitos, in order to detect filarial nematode presence and identify blood-meal sources, respectively. When the parasite genome was detected, the amplicon (cox1 gene, 650 bp fragment) was sent for Sanger sequencing, further confirming the presence of nematodes and species assignation. D. immitis was detected in three Culex pipiens mosquitoes collected in Zrenjanin (August 2021) and Glogonj and Svetozar Miletić (both in July 2021). Additionally, Setaria tundra was detected in Aedes vexans collected in Iđoš (mid-August 2021) and Aedes caspius, which was collected in Mali Iđoš (end of July 2021). This work identifies two new locations where D. immitis occurs in Vojvodina, and is the first report of S. tundra in Serbian territory. Blood-meal analysis provided insights into the preferences of mosquitoes that were positive for Dirofilaria sp. and S. tundra. Full article

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2023

Jump to: 2024, 2022, 2021

12 pages, 1477 KiB

Open AccessArticle

Prevalence and Molecular Characterization of Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi in Diarrheic and Non-Diarrheic Calves from Ningxia, Northwestern China

by Haihui Gao, Gaoxing Liang, Na Su, Qirui Li, Dong Wang, Jiandong Wang, Long Zhao, Xiaodong Kang and Kangkang Guo

Animals 2023, 13(12), 1983; https://doi.org/10.3390/ani13121983 - 14 Jun 2023

Cited by 3 | Viewed by 1604

Abstract

Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi are significant parasitic gastrointestinal pathogens with global distribution in humans and domestic animals, including calves. The main symptoms of calf infection are severe diarrhea, dehydration, growth retardation, and sometimes even death. To date, there has [...] Read more.

Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi are significant parasitic gastrointestinal pathogens with global distribution in humans and domestic animals, including calves. The main symptoms of calf infection are severe diarrhea, dehydration, growth retardation, and sometimes even death. To date, there has been limited information on the prevalence of Cryptosporidium spp., G. duodenalis, and E. bieneusi infections in calves in Ningxia, China, especially between diarrheic and non-diarrheic calves. A total of 438 fecal samples were collected from diarrheic (201) and non-diarrheic (237) calves in Ningxia. PCR and DNA sequencing were used to find the prevalence of Cryptosporidium spp. at 46.8% (205/438), G. duodenalis at 16.9% (74/438), and E. bieneusi at 10.0% (44/438). The prevalence of Cryptosporidium spp. infection in diarrheic and non-diarrheic calves was 54.0% (128/237) and 38.3% (77/201), respectively, and statistical analysis showed a positive correlation between the prevalence of Cryptosporidium spp. infection and calf diarrhea (p < 0.01). However, in this study, there was no statistical correlation between the prevalence of G. duodenalis infection as well as E. bieneusi infection and calf diarrhea (p > 0.05). Furthermore, four known Cryptosporidium species were successfully identified by comparing them with SSU rRNA gene sequences, including C. parvum, C. bovis, C. ryanae, and C. andersoni. In addition, all 74 G. duodenalis-positive samples were identified as assemblage E by comparative analysis of bg gene sequences. Among the 44 E. bieneusi-positive samples sequenced in the present study, 4 distinct E. bieneusi genotypes were successfully identified by comparative analysis of ITS sequences, including 3 known genotypes (J, BEB4, and N) and 1 novel genotype, the latter of which was identified and designated as NX1. These findings indicated that the high genetic diversity and complex population structures of Cryptosporidium spp., G. duodenalis, and E. bieneusi in Ningxia diarrhea calves and non-diarrhea calves, which provide new data for understanding the epidemiological status of Cryptosporidium spp., G. duodenalis, and E. bieneusi in Ningxia calves. Full article

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2022

Jump to: 2024, 2023, 2021

10 pages, 2024 KiB

Open AccessArticle

Development and Preliminary Evaluation of a Nanoparticle-Assisted PCR Assay for the Detection of Cryptosporidium parvum in Calves

by Qian Yao, Xin Yang, Yi Wang, Junwei Wang, Shuang Huang, Junke Song and Guanghui Zhao

Animals 2022, 12(15), 1953; https://doi.org/10.3390/ani12151953 - 1 Aug 2022

Cited by 1 | Viewed by 1807

Abstract

C. parvum is an important diarrheal pathogen in humans and animals, especially in young hosts. To accurately and rapidly detect C. parvum infection in calves, we established a nano-PCR assay targeting the cgd3_330 gene for the specific detection of C. parvum. This [...] Read more.

C. parvum is an important diarrheal pathogen in humans and animals, especially in young hosts. To accurately and rapidly detect C. parvum infection in calves, we established a nano-PCR assay targeting the cgd3_330 gene for the specific detection of C. parvum. This nano-PCR assay was ten times more sensitive than that of the normal PCR assay by applying the same primers and did not cross-react with C. andersoni, C. bovis, C. ryanae, Balantidium coli, Enterocytozoon bieneusi, Giardia lamblia, and Blastocystis sp. To further test the nano-PCR in clinical settings, a total of 20 faecal samples from calves were examined by using the nano-PCR, the normal PCR, and the nested PCR assays. The positive rates were 30% (6/20), 30% (6/20), and 25% (5/20) for the nano-PCR, the normal PCR, and the nested PCR assays, respectively, indicating that the nano-PCR and the normal PCR assays had the same positive rate (30%). Taken together, the present study could provide a candidate method for the specific detection of C. parvum infection in calves in clinical settings. Full article

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12 pages, 9396 KiB

Open AccessArticle

Development of an Immunochromatographic Test Based on Rhoptry Protein 14 for Serological Detection of Toxoplasma gondii Infection in Swine

by Yimin Yang, Yechuan Huang, Xianfeng Zhao, Mi Lin, Lulu Chen, Mingxiu Zhao, Xueqiu Chen, Yi Yang, Guangxu Ma, Chaoqun Yao, Siyang Huang and Aifang Du

Animals 2022, 12(15), 1929; https://doi.org/10.3390/ani12151929 - 28 Jul 2022

Cited by 3 | Viewed by 2231

Abstract

Toxoplasma gondii, a worldwide distributed apicomplexan protozoan, can infect almost all warm-blooded animals and may cause toxoplasmosis. In order to provide a point-of-care detection method for T. gondii infection, an immunochromatographic test (ICT) was established. The proposed test uses recombinant T. gondii [...] Read more.

Toxoplasma gondii, a worldwide distributed apicomplexan protozoan, can infect almost all warm-blooded animals and may cause toxoplasmosis. In order to provide a point-of-care detection method for T. gondii infection, an immunochromatographic test (ICT) was established. The proposed test uses recombinant T. gondii rhoptry protein 14 (ROP14) conjugated with 20 nm gold particles, recombinant protein A as the detection line and monoclonal antibody TgROP14-5D5 as the control line. The specificity, sensitivity, positive predictive value, negative predictive value and stability of this new ICT were evaluated. rTgROP14 was specifically recognized by positive serum of T. gondii but not negative serum. mAb TgROP14-5D5 showed higher specific recognition of T. gondii antigens and was therefore selected for subsequent colloidal gold strip construction. The new ICT based on TgROP14 exhibited good diagnostic performance with high specificity (86.9%) and sensitivity (90.9%) using IHA as a “reference standard”. Among 436 field porcine sera, ICT and IHA detected 134 (30.7%) and 99 (22.7%) positive samples, respectively. The relative agreement was 87.8%. These data indicate that this new ICT based on TgROP14 is a suitable candidate for routine testing of T. gondii in the field. Full article

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14 pages, 778 KiB

Open AccessArticle

Molecular Characterization of Entamoeba spp. in Pigs with Diarrhea in Southern China

by Pei Wang, Sen Li, Yang Zou, Ru-Yi Han, Ping Wang, De-Ping Song, Cheng-Bin Wang and Xiao-Qing Chen

Animals 2022, 12(14), 1764; https://doi.org/10.3390/ani12141764 - 9 Jul 2022

Viewed by 2214

Abstract

Entamoeba spp. is a common zoonotic intestinal protozoan that can parasitize most vertebrates, including humans and pigs, causing severe intestinal diseases and posing a serious threat to public health. However, the available data on Entamoeba spp. infection in pigs are relatively limited in [...] Read more.

Entamoeba spp. is a common zoonotic intestinal protozoan that can parasitize most vertebrates, including humans and pigs, causing severe intestinal diseases and posing a serious threat to public health. However, the available data on Entamoeba spp. infection in pigs are relatively limited in China. To characterize the infection of Entamoeba spp. within pigs in southern China, 1254 fecal samples of diarrheic pigs were collected from 37 intensive pig farms in Hunan, Jiangxi and Fujian provinces and the infection of Entamoeba spp. was investigated based on the small subunit rRNA (SSU rRNA) gene. The overall infection rate of Entamoeba spp. was 58.4% (732/1254), including 38.4% (118/307) in suckling piglets, 51.2% (153/299) in weaned piglets, 57.9% (55/95) in fattening pigs and 73.4% (406/553) in sows, respectively. Moreover, age and the sampling cities in Jiangxi and Fujian provinces were found to be the key factors influencing the infection of Entamoeba spp. (p < 0.05). Two subtypes (ST1 and ST3) with a zoonotic potential of Entamoeba polecki and Entamoeba suis were detected in all age groups of pigs and all sampling areas, with the predominant species and predominant subtype being E. polecki (91.3%, 668/732) and E. polecki ST1 (573/668), respectively, and E. polecki ST1 + E. polecki ST3 (78.6%, 239/304) being the most frequently detected form of mixed infection. Severe Entamoeba spp. infection and zoonotic subtypes were found in this study, exposing a large public health problem in the study area, and strategies need to be implemented to eliminate the risk in the future. Full article

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16 pages, 1668 KiB

Open AccessReview

Epidemiology, Diagnosis, and Prevention of Sparganosis in Asia

by Wei Liu, Tengfang Gong, Shuyu Chen, Quan Liu, Haoying Zhou, Junlin He, Yong Wu, Fen Li and Yisong Liu

Animals 2022, 12(12), 1578; https://doi.org/10.3390/ani12121578 - 18 Jun 2022

Cited by 15 | Viewed by 7005

Abstract

Sparganosis is a zoonotic parasitic disease caused by the larvae (spargana) of the genus Spirometra, which is widely distributed globally and threatens human health. More than 60 species of Spirometra have already been identified, and over 2000 cases have been reported. This [...] Read more.

Sparganosis is a zoonotic parasitic disease caused by the larvae (spargana) of the genus Spirometra, which is widely distributed globally and threatens human health. More than 60 species of Spirometra have already been identified, and over 2000 cases have been reported. This review summarizes the prevalence of humans, frogs, snakes, and other animals with spargana. Furthermore, the infection mode, distribution, and site are summarized and analyzed. We also describe the epidemiology, molecular diagnosis, and other aspects which are of considerable significance to preventing sparganum. Full article

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8 pages, 665 KiB

Open AccessCommunication

The Mitochondrial Genome of Cylicocyclus elongatus (Strongylida: Strongylidae) and Its Comparative Analysis with Other Cylicocyclus Species

by Yuan Gao, Zhonghuai Zhang, Chunren Wang and Kai Zhao

Animals 2022, 12(12), 1571; https://doi.org/10.3390/ani12121571 - 17 Jun 2022

Cited by 2 | Viewed by 1888

Abstract

Cylicocyclus elongatus (C. elongatus) is one of the species in Cylicocyclus, subfamily Cyathostominae, but its taxonomic status in Cylicocyclus is controversial. Mitochondrial (mt) genome is an excellent gene marker which could be used to address the taxonomy controversy. In the [...] Read more.

Cylicocyclus elongatus (C. elongatus) is one of the species in Cylicocyclus, subfamily Cyathostominae, but its taxonomic status in Cylicocyclus is controversial. Mitochondrial (mt) genome is an excellent gene marker which could be used to address the taxonomy controversy. In the present study, the complete mt genome of C. elongatus was determined, and sequence and phylogenetic analyses were performed based on mtDNA data to determine the classification of C. elongatus. The circular complete mt genome of C.elongatus was 13875 bp in size, containing 12 protein-coding genes (12 PCGs), 22 transfer RNA (tRNA) genes, 2 ribosomal RNA (rRNA) genes, and 2 non-coding regions (NCRs). The A + T content of C. elongatus complete mt genome was 76.64%. There were 19 intergenic spacers with lengths of 2–53 bp and 2 overlaps with lengths of 1–2 bp in the impact complete mt genome. ATT and TAA were the most common start and termination codons of 12 PCGs, respectively. Comparative analyses of mt genomes nucleotide sequence and amino acid sequence showed that there were higher identities between C. elongatus and five other Cylicocyclus, rather than with P. imparidentatum. Phylogenetic analyses based on concatenated nucleotide sequences of 12 PCGs of 23 species in the family Strongylidae showed that C. elongatus was closely related to Cylicocyclus species, rather than P. imparidentatum. We concluded that C. elongatus was a member in Cylicocyclus based on comparative and phylogenetic analyses of mt genome sequences. The data of the complete mt genome sequence of C. elongatus provide a new and useful genetic marker for further research on Cyathostominae nematodes. Full article

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15 pages, 1603 KiB

Open AccessArticle

Molecular Characterization of Anaplasma spp. among Dairy, Cashmere, and Meat Goats in Shaanxi Province, Northwestern China

by Xin Yang, Mingzhe Fu, Zhengqing Yu, Junwei Wang, Junke Song and Guanghui Zhao

Animals 2022, 12(12), 1566; https://doi.org/10.3390/ani12121566 - 17 Jun 2022

Cited by 5 | Viewed by 1893

Abstract

Anaplasma spp. are important tick-borne pathogens endangering the health of humans and various animals. Although several studies have reported Anaplasma infection in livestock in China, little is known about the impact of production categories on the occurrence of Anaplasma species. In the present [...] Read more.

Anaplasma spp. are important tick-borne pathogens endangering the health of humans and various animals. Although several studies have reported Anaplasma infection in livestock in China, little is known about the impact of production categories on the occurrence of Anaplasma species. In the present study, PCR tools targeting the 16S rRNA and msp4 genes were applied to investigate the prevalence of Anaplasma spp. in 509 blood samples of dairy (n = 249), cashmere (n = 139), and meat (n = 121) goats from Shaanxi province. The prevalence of Anaplasma spp. was 58.5% (298/509) in goats, and significant differences (p < 0.001) were identified in the prevalence among production categories, with the highest in meat goats (84.3%, 102/121), followed by cashmere goats (58.3%, 81/139) and dairy goats (46.2%, 115/249). Significant differences (p < 0.001) in prevalence were also found among sampling sites and age groups. Meanwhile, the prevalence was 36.9% (188/509) for A. phagocytophilum, 36.1% (184/509) for A. bovis, and 11.0% (56/509) for A. ovis, and significant differences (p < 0.001) in prevalence of A. phagocytophilum, A. bovis and A. ovis were recognized among production categories and sampling sites. A. phagocytophilum, A. bovis and A. ovis were dominant species in meat, dairy, and cashmere goats, respectively, and A. ovis was absent in meat goats. Co-infections were found in 124 (24.4%) investigated samples. Goats aged < 2, 3–6, and 7–12 months, and goats from Qingjian and Zhenba were risk factors associated with the occurrence of Anaplasma. Phylogenetic analysis indicated separate clades for the distribution of A. phagocytophilum from different ruminant, reflecting potential host adaption within this species. This study reported the colonization occurrence of Anaplasma spp. among production categories in goats in Shaanxi province and enriched our knowledge on the transmission of Anaplasma spp. in goats in China. Considering the existence of zoonotic A. phagocytophilum in goats in this study and previous reports, interventions based on One Health are needed to be developed to control the transmission of Anaplasma spp. between humans and animals. Full article

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10 pages, 2059 KiB

Open AccessArticle

Molecular Characterization of 18S rDNA, ITS-1, ITS-2, and COI from Eimeria christenseni and E. arloingi in Goats from Shaanxi Province, Northwestern China

by Gaoxing Liang, Xin Yang, Ding Liu, Yuan Li, Junwei Wang, Xi Chen, Guanghui Zhao and Junke Song

Animals 2022, 12(11), 1340; https://doi.org/10.3390/ani12111340 - 24 May 2022

Viewed by 3667

Abstract

Coccidiosis caused by Eimeria is one of the most common and significant diseases in goats, leading to serious economic losses in the development of the goat industry. Although several genetic loci, such as 18S rDNA, ITS-1, ITS-2, and COI, have been applied in [...] Read more.

Coccidiosis caused by Eimeria is one of the most common and significant diseases in goats, leading to serious economic losses in the development of the goat industry. Although several genetic loci, such as 18S rDNA, ITS-1, ITS-2, and COI, have been applied in the molecular characterization of Eimeria in chicken, rabbits, turkey, and wildlife, little is known about these molecular markers of Eimeria in goats. In the present study, we isolated purified oocysts of highly pathogenic Eimeriaarloingi and Eimeria christenseni from fecal samples of goats in Shaanxi province, China, and then subjected these purified oocysts to genomic DNA isolation, PCR amplification, and sequencing of 18S rDNA, ITS-1, ITS-2, and COI loci of Eimeria arloingi and Eimeria christenseni. Finally, the obtained sequences were used for phylogenetic analysis of Eimeria species in goats and other livestock. The lengths of the 18S rDNA, ITS-1, ITS-2, and COI were 1790 bp, 403 bp, 584 bp, and 1268 bp for E. arloingi and 1796 bp, 386 bp, 565 bp, and 1268 bp for E. christenseni, respectively. The phylogenetical analysis based on 18S rDNA indicated that E. christenseni and E. arloingi were the most closely related to ovine Eimeria, followed by E. bovis, E. ellipsoidalis, and E. zuernii from cattle. The phylogenetical analysis based on ITS-1 and ITS-2 could not effectively distinguish ovine Eimeria from caprine Eimeria. The phylogenetical analysis based on the COI locus could effectively distinguish between Eimeria species from goats and cattle, but it was ineffective in distinguishing between Eimeria species from sheep and goats. To the best of our knowledge, this is the first characterization of 18S rDNA, ITS-1, ITS-2, and COI in E. arloingi and E. christenseni; it can provide useful genetic markers for molecular epidemiological and population genetic studies on E. arloingi and E. christenseni in goats and contribute to the prevention and control of goat coccidiosis. Full article

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11 pages, 1538 KiB

Open AccessArticle

Epidemiology and Genetic Diversity of Spirometra Tapeworm Isolates from Snakes in Hunan Province, China

by Tengfang Gong, Xiaoyi Su, Fen Li, Junlin He, Shuyu Chen, Wenchao Li, Xinrui Xie, Yisong Liu, Xi Zhang and Wei Liu

Animals 2022, 12(9), 1216; https://doi.org/10.3390/ani12091216 - 9 May 2022

Cited by 2 | Viewed by 1931

Abstract

Sparganosis, caused by the plerocercoid larvae of Spirometra tapeworms, is a public health hazard worldwide. The prevalence and genetics of sparganum from snakes remain unclear. In this study, we investigated the prevalence of sparganum infection in wild snakes in Hunan province and compared [...] Read more.

Sparganosis, caused by the plerocercoid larvae of Spirometra tapeworms, is a public health hazard worldwide. The prevalence and genetics of sparganum from snakes remain unclear. In this study, we investigated the prevalence of sparganum infection in wild snakes in Hunan province and compared the prevalence of Spirometra tapeworms in snakes worldwide. Furthermore, the genetic diversity of collected isolates was analyzed using mitochondrial cytb and cox1 genes. The result shows that the sparganum infection rate in wild snakes (89.50%, 402/449) was higher in Hunan than in other regions. Genetic diversity analysis based on concatenated sequences revealed high genetic diversity but no distinct genetic structure among Spirometra populations. Phylogenetic analysis supported the division of European and Chinese Spirometra isolates and a single species in Chinese Spirometra isolates. The prevalence of Spirometra tapeworms in snakes is serious, and the risk of sparganosis should be further publicized. Full article

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15 pages, 1808 KiB

Open AccessFeature PaperArticle

Establishment and Application of an Indirect Enzyme-Linked Immunosorbent Assay for Measuring GPI-Anchored Protein 52 (P52) Antibodies in Babesia gibsoni-Infected Dogs

by Qin Liu, Xueyan Zhan, Dongfang Li, Junlong Zhao, Haiyong Wei, Heba Alzan and Lan He

Animals 2022, 12(9), 1197; https://doi.org/10.3390/ani12091197 - 6 May 2022

Cited by 1 | Viewed by 2053

Abstract

Babesia gibsoni is a malaria-like protozoan that parasitizes the red blood cells of canids to cause babesiosis. Due to its high expression and essential function in the survival of parasites, the Glycosylphosphatidylinositol (GPI) anchor protein family is considered an excellent immunodiagnostic marker. Herein, [...] Read more.

Babesia gibsoni is a malaria-like protozoan that parasitizes the red blood cells of canids to cause babesiosis. Due to its high expression and essential function in the survival of parasites, the Glycosylphosphatidylinositol (GPI) anchor protein family is considered an excellent immunodiagnostic marker. Herein, we identified a novel GPI-anchored protein named as BgGPI52-WH with a size of 52 kDa; the recombinant BgGPI52-WH with high antigenicity and immunogenicity was used as a diagnostic antigen to establish a new iELISA method. The iELISA had a sensitivity of 1:400, and no cross-reaction with other apicomplexan parasites occurred. We further demonstrated that the degree of variation was less than 10% using the same samples from the same or different batches of an enzyme-labeled strip. It was found that the method was able to detect early infection (6 days after infection) in the sera of the B. gibsoni-infected experimental dogs in which antibody response to rBgGPI52-WH was evaluated. Clinical sera from pet hospitals were further tested, and the average positive rate was about 11.41% (17/149). The results indicate that BgGPI52-WH is a reliable diagnostic antigen, and the new iELISA could be used as a practical method for the early diagnosis of B. gibsoni. Full article

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8 pages, 624 KiB

Open AccessArticle

Molecular Identification and Genotyping of Enterocytozoon bieneusi in Sheep in Shanxi Province, North China

by Rui-Lin Qin, Ya-Ya Liu, Jin-Jin Mei, Yang Zou, Zhen-Huan Zhang, Wen-Bin Zheng, Qing Liu, Wen-Wei Gao, Shi-Chen Xie and Xing-Quan Zhu

Animals 2022, 12(8), 993; https://doi.org/10.3390/ani12080993 - 12 Apr 2022

Cited by 3 | Viewed by 2042

Abstract

Enterocytozoon bieneusi is a fungus-like protist that can cause malabsorption and diarrhea in sheep, other animals, and humans, threatening the development of animal husbandry and public health. To date, there are no data about the prevalence and genotypes of E. bieneusi in sheep [...] Read more.

Enterocytozoon bieneusi is a fungus-like protist that can cause malabsorption and diarrhea in sheep, other animals, and humans, threatening the development of animal husbandry and public health. To date, there are no data about the prevalence and genotypes of E. bieneusi in sheep in Shanxi Province, North China. In this study, 492 fecal samples were collected from sheep in three representative counties in northern, central, and southern Shanxi Province. Nested PCR amplification was performed to detect the prevalence and identify the genotypes of E. bieneusi based on the internal transcribed spacer (ITS) region of the rRNA gene. Overall, 168 of 492 examined samples were E. bieneusi-positive, with a prevalence of 34.2% (168/492). Significant differences in the prevalence of E. bieneusi were observed among the three sampled regions (χ2 = 95.859, df = 2, p < 0.001), but the differences in E. bieneusi prevalence were not statistically significant between different genders and age groups (p > 0.05). Sequence analysis showed that four known genotypes (BEB6, COS-I, CHS7, and CHC8) and one novel genotype (named SY-1) were identified. BEB6 was the prevalent genotype found within the three counties. Phylogenetic analysis revealed that the five genotypes observed in this study belong to Group 2. The present study reported the presence and genotypes of E. bieneusi infection in sheep in Shanxi Province for the first time, which enriches the knowledge of the genetic diversity of E. bieneusi and provides baseline data for the prevention and control of E. bieneusi infection in animals and humans. Full article

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11 pages, 3330 KiB

Open AccessArticle

Positivity Rate Investigation and Anthelmintic Resistance Analysis of Gastrointestinal Nematodes in Sheep and Cattle in Ordos, China

by Bin Hou, Rong Yong, Jiya Wuen, Yong Zhang, Buhe Buyin, Dihua Subu, Huhen Zha, Hong Li and Surong Hasi

Animals 2022, 12(7), 891; https://doi.org/10.3390/ani12070891 - 31 Mar 2022

Cited by 7 | Viewed by 2720

Abstract

Gastrointestinal nematodes (GINs), such as Trichostrongylidae, are important pathogens in small ruminants, causing significant losses in these livestock species. Despite their veterinary importance, GINs have not been studied in certain regions of the world. Therefore, much of their epidemiology and economic impact [...] Read more.

Gastrointestinal nematodes (GINs), such as Trichostrongylidae, are important pathogens in small ruminants, causing significant losses in these livestock species. Despite their veterinary importance, GINs have not been studied in certain regions of the world. Therefore, much of their epidemiology and economic impact on production remain unknown. In the present study, a systematic epidemiological survey based on the modified McMaster technique was conducted to investigate the type and infection of GINs in sheep and cattle. In 9622 fecal samples from 491 sampling sites in the four main banner districts of Ordos, the prevalence of GIN infection was found to be 38.84% and 4.48% in sheep and cattle, respectively. At the same time, the effects of four pasture types on the distribution of GINs were analyzed. This study also found severe resistance to ivermectin and albendazole in GINs and suspected anthelmintic resistance in nitroxynil, levamisole and closantel. We report the type and infection of GINs in Ordos, with the aim to help the prevention and control of GINs. Based on the results of the questionnaire survey and GIN resistance test, we found several reasons for the anthelmintic resistance of GINs, consequently providing new ideas for controlling the occurrence of anthelmintic resistance. Full article

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18 pages, 3870 KiB

Open AccessArticle

Mutation Characteristics and Phylogenetic Analysis of Five Leishmania Clinical Isolates

by Zhiwan Zheng, Jinlei He, Tao Luo, Jianhui Zhang, Qi Zhou, Shuangshuang Yin, Dali Chen, Jie Luo, Jianping Chen and Jiao Li

Animals 2022, 12(3), 321; https://doi.org/10.3390/ani12030321 - 28 Jan 2022

Cited by 2 | Viewed by 2216

Abstract

Leishmaniasis is a neglected tropical disease threatening millions of people worldwide. The emergence of antimony-resistant Leishmania strains have brought difficulties to the treatment and elimination of leishmaniasis. This study performed genome sequencing, phylogenetic analysis and mutation analysis of five Leishmania clinical isolates, especially [...] Read more.

Leishmaniasis is a neglected tropical disease threatening millions of people worldwide. The emergence of antimony-resistant Leishmania strains have brought difficulties to the treatment and elimination of leishmaniasis. This study performed genome sequencing, phylogenetic analysis and mutation analysis of five Leishmania clinical isolates, especially the Leishmania strain L_HCZ isolated in 2016, which shows strong virulence and antimony resistance. By phylogenetic analysis, four isolates (L_DD8, L_801, L_Liu and L_9044) were identified as Leishmania donovani, the isolate L_HCZ was identified as Leishmania infantum and the isolate L_DD8 as a standard strain of L.donovani. Genome-wide mutation analysis was applied to identify mutations related to the drug resistance and virulence of the newly isolated L_HCZ. Compared with the other four Leishmania isolates, L_HCZ had the most mutations in genes associated with antimony resistance, including the ABC transporter, ascorbate-dependent peroxidase, gamma–glutamylcysteine synthetase, glucose-6-phosphate 1-dehydrogenase, ATP-binding cassette protein subfamily A and multi-drug resistance protein-like genes. Among the genes associated with virulence, L_HCZ had the most mutations in cysteine peptidase A, cysteine peptidase B, cysteine peptidase C, heat-shock protein 70, gp63, acid phosphatase, kinesin k39, kinesin, phosphoglycan beta 1, amastin-like surface protein and amastin-like proteins. The mutations in L_HCZ might possibly contribute to its antimony resistance and strong virulence in clinical patients. Whole-genome resequencing has exhibited broad application prospects and may be put into clinical use in the future for parasite identifying and epidemiological investigations. Full article

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2021

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9 pages, 463 KiB

Open AccessArticle

Genotyping and Zoonotic Potential of Enterocytozoon bieneusi in Stray Dogs Sheltered from Shanghai, China

by Hua Liu, Jie Xu, Yujuan Shen, Jianping Cao and Jianhai Yin

Animals 2021, 11(12), 3571; https://doi.org/10.3390/ani11123571 - 16 Dec 2021

Cited by 3 | Viewed by 2375

Abstract

Microsporidia are considered to be highly diverged and specialized parasites, and can infect a wide variety of vertebrate and invertebrate hosts. Enterocytozoon bieneusi is the most prevalent species in humans and various livestock, wildlife, and companion mammals. Dogs being the most popular companion [...] Read more.

Microsporidia are considered to be highly diverged and specialized parasites, and can infect a wide variety of vertebrate and invertebrate hosts. Enterocytozoon bieneusi is the most prevalent species in humans and various livestock, wildlife, and companion mammals. Dogs being the most popular companion animals of humans become more and more regarded. In this study, 272 fecal specimens were collected from stray dogs from Shanghai, but the dogs were adopted in a shelter in Zhenjiang, Jiangsu Province, China. E. bieneusi was examined by PCR amplification of the internal transcribed spacer (ITS) region and sequence analysis. The total positive rate of E. bieneusi was 8.8% (24/272). Moreover, 8 genotypes were found, including three known (genotypes EbpA, Henan V and Type IV) and 5 novel genotypes (genotypes SHZJD1–5). Two samples were positive for two genotypes, one was positive fortype SHZJD4 and Henan V, the other was positive for Henan V and Type IV. In addition, phylogenetic analysis showed all genotypes obtained in this study were all clustered into the zoonotic group 1. Therefore, the risk of zoonotic transmission of pathogens such as E. bieneusi from stray dogs to humans potentially threaten human health, and it is time to strengthen their health management. Full article

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9 pages, 1001 KiB

Open AccessCommunication

Molecular Characterization of Blastocystis sp. in Camelus bactrianus in Northwestern China

by Xin Yang, Yunhui Li, Yuxin Wang, Junwei Wang, Peng Lai, Yuan Li, Junke Song, Meng Qi and Guanghui Zhao

Animals 2021, 11(11), 3016; https://doi.org/10.3390/ani11113016 - 20 Oct 2021

Cited by 12 | Viewed by 1746

Abstract

Blastocystis sp. is an important zoonotic protist in humans and various animals with worldwide distribution. However, there have been no data on the occurrence of Blastocystis sp. in C. bactrianus, an important economic animal in northwestern China. In the present study, a [...] Read more.

Blastocystis sp. is an important zoonotic protist in humans and various animals with worldwide distribution. However, there have been no data on the occurrence of Blastocystis sp. in C. bactrianus, an important economic animal in northwestern China. In the present study, a PCR-sequencing tool based on the SSU rRNA gene was applied to investigate the prevalence and genetic diversity of Blastocystis sp. in 638 faecal samples from C. bactrianus in 21 sampling sites within three main breeding areas (Gansu, Inner Mongolia and Xinjiang) in northwestern China. The total prevalence of Blastocystis sp. was 21.8% (139/638) in C. bactrianus, with the infection rates of 29.5% (18/61), 50.0% (14/28) and 19.5% (107/549) for animals aged <2 years, 2–6 years and >6 years, respectively. Significant differences in prevalence were detected among C. bactrianus from three geographic areas (χ² = 19.972, df = 2, p < 0.001) and all sampling sites (χ² = 104.154, df = 20, p < 0.001). A total of 16 of 21 sampling sites were positive for Blastocystis sp., with the prevalence ranging from 7.7% to 70.6%. Sequence analysis of the SSU rRNA gene identified eight subtypes in C. bactrianus in the present study, including seven animal adapted subtypes (ST10, ST14, ST21, ST24, ST25, ST26 and ST30) and one potentially novel subtype, with ST10 being the dominant one. To the best of our knowledge, this study provides the first insight for the occurrence and genetic make-up of Blastocystis sp. in C. bactrianus and contributes to the understanding of the transmission of Blastocystis infection in C. bactrianus in China. Full article

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9 pages, 11680 KiB

Open AccessCommunication

Prevalence and Novel Genotypes Identification of Enterocytozoon bieneusi in Dairy Cattle in Yunnan Province, China

by Hai-Yang Song, Kai-Sheng Wang, Jian-Fa Yang, Hua-Ming Mao, Li-Hua Pu, Yang Zou, Jun Ma, Xing-Quan Zhu, Feng-Cai Zou and Jun-Jun He

Animals 2021, 11(11), 3014; https://doi.org/10.3390/ani11113014 - 20 Oct 2021

Cited by 6 | Viewed by 1691

Abstract

Enterocytozoon bieneusi is a fungus-like protist parasite that can cause diarrhea and enteric diseases. The infection of E. bieneusi has been reported in many host species, including cattle and humans. However, information on prevalence and genotype distribution of E. bieneusi in dairy cattle [...] Read more.

Enterocytozoon bieneusi is a fungus-like protist parasite that can cause diarrhea and enteric diseases. The infection of E. bieneusi has been reported in many host species, including cattle and humans. However, information on prevalence and genotype distribution of E. bieneusi in dairy cattle in Yunnan province in China is still absent. In this study, 490 Holstein Cows and 351 dairy buffalo fecal samples were collected from three regions in Yunnan province, China. By using nest-PCR that targets the internal transcribed spacer (ITS), we found that the prevalence of E. bieneusi was 0.59% (5/841). DNA sequence analysis showed that five E. bieneusi genotypes were identified in this study, including two novel genotypes, YNDCEB-90 and YNDCEB-174, and three known genotypes (I, J, BEB4). Phylogenetic analysis revealed that two novel genotypes, YNDCEB-90 and YNDCEB-174, were clustered into Group 1, representing the zoonotic potential. The remaining genotypes I, J, and BEB4, which are the most frequent genotypes of E. bieneusi infection in cattle and lead to E. bieneusi infection in humans, belonged to Group 2. Although the lower prevalence of E. bieneusi was detected in dairy cattle in Yunnan province, it indicates that dairy cattle should be considered to be one of the potential hosts for transmitting E. bieneusi to humans. These findings are important for the development of effective prevention strategies for microsporidiosis. Full article

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