Donkeys In China Research Articles

Prevalence and Multilocus Genotyping of Giardia duodenalis in Donkeys in Shanxi Province, North China.

Giardia duodenalis is a ubiquitous flagellated protozoan, causing significant economic losses to animal husbandry and posing threats to public health. China ranks the world's sixth largest major producer of donkeys, rearing approximately 2.6 million donkeys in 2019, but limited investigation of G. duodenalis prevalence has been conducted in the past, and it is yet to be known whether donkeys in Shanxi Province are infected with G. duodenalis. In the present study, a total of 815 fecal samples collected from donkeys in representative regions of Shanxi Province, North China, were examined for G. duodenalis using nested PCR. Then, the assemblages and multilocus genotypes (MLGs) were examined based on three established loci: namely, β-giardin (bg), triosephosphate isomerase (tpi), and glutamate dehydrogenase (gdh). The overall prevalence of G. duodenalis in donkeys in Shanxi Province was 16.81% (137/815). The region was identified as the main risk factor for the observed difference in G. duodenalis prevalence in donkeys among the three study areas (χ2 = 21.611, p < 0.001). Assemblages A, E, and B were identified, with the latter as the predominant assemblage. Three MLGs (MLG-novel-1 to 3) were formed based on sequence variation among the three loci. The present study reveals the presence of G. duodenalis in donkeys in Shanxi Province, North China, for the first time, which not only enriches the data on the distribution of G. duodenalis in donkeys in China but also provides useful baseline data for planning control strategies against G. duodenalis infection in the sampled areas.

Characteristics and epidemiological investigation of equid herpesvirus 8 in donkeys in Shandong, China.

Equid herpesvirus 8 (EHV-8), also known as asinine herpesvirus type 3 (AHV-3), can cause severe respiratory disease, abortion in mares, and neurological disorders. There is limited information on the prevalence of EHV-8 in donkeys in China. In this study, we investigated EHV-8 infection in donkeys using PCR, resulting in the identification of a field strain, termed EHV-8 SD2020113, which was isolated using RK-13 cells and characterized by high-throughput sequencing and transmission electron microscopy. Our data indicated that 38.7% (457/1180) of donkeys showed the presence of EHV-8 in blood samples. Analysis of the ORF70 gene showed the highest similarity (99.8-99.9% identity) to EHV-8 IR/2015/40 (MF431614.1) and SDLC66 (MW816102), and, in phylogenetic analysis, it clustered with EHV-8 SDLC66 from China. The findings of this study indicate that EHV-8 is likely to represent a threat to the donkey industry, and breeders andveterinarians who care for donkey farms should be aware of this.

Novel Streptococcus equi strains causing strangles outbreaks in Arabian horses in Egypt.

Strangles displays a major challenge to veterinary medicine worldwide. However, no data on Streptococcus equi subsp. equi (S. equi) M protein alleles have been reported so far from Arabian horses. We report here for the first time the S. equi SeM alleles causing strangles in Arabian horses, and the associated risk factors for the disease. Duplicate samples from one hundred Arabian horses with acute strangles in confirmed outbreaks and sporadic cases were analysed by phenotypic methods and multiplex polymerase chain reaction (PCR) targeting streptokinase precursor, seeI and sodA genes. PCR and sequencing of S. equi SeM gene were employed for strains typing, and the four superantigens were determined among the allelic variants. Direct-sample PCR confirmed and highly positively correlated (r=.85) with the phenotypic results, and detected S. equi in five samples more than the conventional culture. A combination of multiplex PCR from samples and culture could successfully identify S. equi (92%), S. zooepidemicus (5%) and S. equisimilis (3%). SeM typing demonstrated five SeM alleles, including four previously unidentified alleles that were deposited in the PubMLST-SeM database. SeM-139 and SeM-141 are related to some strains that were recently recovered from donkeys in China. SeM-140 and SeM-199 are related to a group of alleles from horses in Europe. Variation in the presence of seeM, seeH and seeL superantigens was found across the four novel alleles without interference with the severity of strangles and clinical presentation seen in different outbreaks. Horse age was the most important factor in developing strangles, followed by seasonality and the diagnosis of strangles in the previous year. These new findings comprise a significant contribution to the horse industry through the identification of novel S. equi SeM types that may bolster measures for strangles control as the identified SeM alleles will certainly help in the development of SeM-containing vaccine.

An outbreak of strangles associated with a novel genotype of Streptococcus equi subspecies equi in donkeys in China during 2018.

Strangles is a highly contagious respiratory disease of equids caused by the bacterium Streptococcus equi subspecies equi. To identify the cause of an outbreak of strangles that occurred on donkey farms within the Shandong Province of China and determine the prevalence of the disease. Cross-sectional. Samples were taken from clinically affected animals to measure the prevalence of strangles within the population of donkeys at six intensive farms in China and identify the SeM type of isolate recovered from affected animals. Diagnosis was confirmed by bacterial isolation, biochemical tests and PCR. Epidemiological data were analysed using Chi-square test and a Fisher's exact two-sided test. The SeM gene of S.equi isolates recovered from affected animals was determined and compared with the SeM database PubMLST-seM. In July and August 2018, an outbreak of strangles occurred on six donkey farms within the Shandong Province of China. The overall prevalence of disease within the different donkey herds was 13.4%. Younger animals were worst affected with 40.3% (83/206) of donkey foals aged under 1year exhibiting clinical signs compared with 12.5% (191/1525) of donkeys aged one to 2years and 3.8% (17/442) of donkeys over 2years of age. Analysis of SeM sequencing data identified that the farms were affected by the same strain of S.equi, SD201807, which contains the novel 136 allele of SeM. Healthy donkeys were not sampled in this study. The number of intensive donkey breeding farms in China has risen recently. The higher numbers of animals that are in closer proximity to one another raise the potential for the transmission of infectious diseases such as strangles. This is the first description of a strangles outbreak among donkey herds in China. The Summary is available in Chinese - see Supporting information.

First genetic characterization of Toxoplasma gondii infection in donkey meat slaughtered for human consumption in Shandong province, eastern China

Until now, limited information on the molecular epidemiology and genotypes of Toxoplasma gondii infection in donkeys is available in China. Thus, the present study was conducted to characterize T. gondii genotypes in donkeys, intended for human consumption in Shandong province, eastern China. A total of 618 muscle tissue samples of donkeys was collected in Shandong province from January 2016 to August 2017 and were used to detect the T. gondii B1 gene by a semi-nested PCR, and the positive samples were genotyped at 10 nuclear loci (i.e., SAG1, alternative SAG2, 5′-and 3′-SAG2, SAG3, L358, BTUB, c22-8, GRA6, c29-2, PK1) and an apicoplast locus Apico by multi-locus PCR-RFLP method. Fifty-seven (9.22%) samples out of 618 donkey meat samples were examined T. gondii B1 gene positive. In this study, no risk factor was found to be associated with T. gondii infection in donkeys. Moreover, two genotypes (ToxoDB#9 and ToxoDB#1) were identified. This is the first genetic characterization of T. gondii isolated from donkey meat that would intend for human consumption in Shandong province, eastern China and also the first report of genotype ToxoDB#1 found in donkeys in China, which may be useful for preventing and controlling T. gondii infection in donkeys, other animals and humans.

Morphological and molecular characteristics of Sarcocystis bertrami from horses and donkeys in China

While Sarcocystis parasites from the muscles of donkey and horse have been characterized as different species, similarities between the parasites in these host raises questions about this assignment (Levine and Tadros, 1980; Matuschka, 1983; Odening et al., 1995b). To resolve this, we examined the tissue cysts of Sarcocystis collected from donkeys and horses were studied by morphological and molecular methods. Morphological studies performed by light microscopy (LM) revealed that each of two types of cysts were present in samples from each host type. Under LM, villar protrusions (VP) were sometimes observed on the larger (Type I) and smaller (Type II) of these cyst types; when present, these were sometimes short and sometimes long. By electron microscopy (EM), VPs from both horse and donkey cysts were found to share similar structures, appearing to be typical of ‘type 11a’ VPs found on the Sarcocystis wall of Sarcocystis fayeri as described by Dubey et al., 1977. The VP of cysts in both horses and donkeys contained microtubules extending from the villar tips to the ground substance (GS). Ovoid, osmiophilic bodies (OB) were found along the length of the microtubules within the villi, but this feature was not found in all VP. To understand the phylogeny of the parasites, a portion of the coxI gene was sequenced from 22 isolated cysts (9 from donkeys and 13 from horses). Phylogenetic relationships were reconstructed from these sequences and the closest homologues available in GenBank, revealing that all of the samples, regardless of host origin or morphological appearance under LM, grouped in one clade. Ours is the first attempt to combine morphological measurements with coxI sequences in assessing such equine parasites; the results confirm a close relationship of the parasites from horse and donkey with S. fayeri. Further, the data suggest that the cysts in each host likely belong to the same species. As the first named species was Sarcocystis bertrami, we propose S. bertrami (syn. Sarcocystis fayeri) as the descriptor for this parasite of both horses and donkeys. Ultimately, this finding will only be validated by cross-transmission infection experiments that score the ability of parasite isolates from one Equus to infect the other.

Seroprevalence of Toxoplasma gondii infection and variables associated with seropositivity in donkeys in eastern China.

Donkeys (Equus asinus) are widely distributed throughout China; they are used for their meat, as food, and certain donkey-derived items are also important for traditional Chinese medicinal purposes. However, only limited information is available on Toxoplasma gondii infection in donkeys in China, especially the eastern region, which is one of the largest production areas. Thus, the present study was conducted to detect specific anti-T. gondii antibodies using a commercially available indirect hemagglutination test (IHA) kit and to evaluate the risk factors that are associated with seroprevalence in the Shandong province of eastern China. A total of 213/1278 (17%) donkeys tested from Shandong province were positive for T. gondii antibodies. Statistical analysis revealed that gender and feeding habits of the animal are associated with T. gondii infection. These results provide information for the prevention and control of toxoplasmosis in donkeys, other animals, and humans in this region and elsewhere.

Prevalence of Neospora spp. in donkeys in China.

This study was conducted to detect specific anti-Neospora antibodies using a commercial competitive-inhibition ELISA kit, and to evaluate the risk factors for Neospora spp. infection. Out of a total of 2,228 donkey sera collected in three provinces in China, 211 (9.5%) were found to be positive for anti-Neospora antibodies. Statistical analysis revealed that age (p = 0.019, OR = 1.62, 95%CI: 1.08-2.44), feeding status (p < 0.001, OR = 3.79, 95%CI: 2.65-5.43), miscarriage history (p = 0.006, OR = 2.56, 95%CI: 1.27-4.01), and contact with dogs (p < 0.001, OR = 2.69, 95%CI: 1.86-3.88) were significant risk factors for Neospora spp. infection. This is the first evidence of Neospora infection in donkeys in China.

Emergence of H3N8 equine influenza virus in donkeys in China in 2017

Equine influenza virus is a major respiratory pathogen in horses. Although both horses and donkeys belong to the genus Equus, donkey infection with influenza viruses is rare. In March 2017, an influenza outbreak occurred in donkeys in Shandong province, China. The causative virus, A/donkey/Shandong/1/2017(H3N8), was isolated from a dead donkey. Genetic analysis indicated that the virus originated from influenza A (H3N8) clade 2 of the Florida sub-lineage that has been circulating in Asian equine populations. Comparison of the deduced amino acid sequence of the HA gene of this causative virus with that of the A/equine/Richmond/1/2007 vaccine strain showed that substitutions had occurred in the antigenic regions A, B, and C. This study provides insight into the currently circulating and newly emerging H3N8 strains in donkeys in China.

Occurrence of Enterocytozoon bieneusi in Donkeys (Equus asinus) in China: A Public Health Concern.

Enterocytozoon bieneusi is an important zoonotic parasite. It can infect virtually all animal species and has a global distribution. However, the prevalence of E. bieneusi in donkeys (Equus asinus) has only been reported in Algeria and Spain, and no information is available concerning genotypes of E. bieneusi in donkeys worldwide. In the present study, a total of 301 donkey fecal samples (48 from Jilin Province, 224 from Shandong Province and 29 from Liaoning Province) were collected and examined by PCR amplification of the internal transcribed spacer (ITS) region. The overall E. bieneusi prevalence was 5.3% (16/301), with 6.3% (3/48) in Jilin Province, 4.9% (11/224) in Shandong Province, and 6.9% (2/29) in Liaoning Province. Prevalence in different age groups ranged from 4.2 to 5.5%. E. bieneusi prevalence in donkeys sampled in different seasons varied from 4.2 to 6.5%. Altogether, four E. bieneusi genotypes were identified in this study, with two known genotypes (J and D) and two novel genotypes (NCD-1and NCD-2). Phylogenetic analysis revealed that genotypes D, NCD-1 and NCD-2 belonged to group 1, while the remaining genotype J was clustered into group 2. These findings revealed the occurrence of E. bieneusi in donkeys in China for the first time. Moreover, the present study also firstly genotyped the E. bieneusi in donkeys worldwide. These findings extend the distribution of E. bieneusi genotypes and provide baseline data for controlling E. bieneusi infection in donkeys, other animals and humans.

The presence of Giardia intestinalis in donkeys, Equus asinus, in China

BackgroundGiardia intestinalis is one of the most important zoonotic enteric parasites. As no information regarding prevalence and genotype of G. intestinalis in donkeys (Equus asinus) in China is available, 181 faecal samples from 48 donkeys from Jilin Province, from 104 from Shandong Province and from 29 from Liaoning Province were examined between May and December 2015.FindingsTwenty-eight (15.47%) out of 181 donkey samples were tested G. intestinalis-positive by nested amplification of the triosephosphate isomerase (tpi) gene. The prevalence in different regional groups varied from 10.42 to 18.27%. The prevalence in adult and young donkeys was 14.29 and 22.92%, respectively. Otherwise, the prevalence was 11.69% in summer and 18.27% in winter. However, no statistically significant differences were found in relation to region or age group. Sequence analysis of the tpi, glutamate dehydrogenase (gdh) and beta giardin (bg) loci identified 4, 1 and 3 subtypes of assemblage B, respectively. Moreover, four novel multilocus genotypes (MLGs novel-1 to novel-4) were identified in assemblage B.ConclusionsThis first report of G. intestinalis in donkeys in China indicates that further studies of nation-wide molecular epidemiology and geographical distribution of Giardia in donkeys are warranted. Effective strategies should be implemented to control G. intestinalis infection in donkeys, other animals and humans.

Seroprevalence of Toxoplasma gondii in slaughtered horses and donkeys in Liaoning province, northeastern China

BackgroundToxoplasma gondii is an important zoonotic pathogen infecting humans and almost all warm-blooded animals. The most common sources of human infection are ingestion of tissue cysts in raw or undercooked meat. However, limited information is available about T. gondii infection in horses and donkeys in China. In the present study, we report the seroprevalence of T. gondii infection in horses and donkeys in Liaoning province, northeastern China.MethodsSerum samples were collected from 711 and 738 slaughtered horses and donkeys, respectively, in 13 regions of Liaoning province. The modified agglutination test (MAT) was used to test the specific antibodies to T. gondii.ResultsIn this study, 178 of 711 (25.0%) horses were seropositive for T. gondii with titers of 1:25 in 81, 1:50 in 62, 1:100 in 33, and 1:200 in 2, and seroprevalence of T. gondii infection from 13 regions ranged from 8.2% to 37.0%. Antibodies to T. gondii were found in 174 of 738 (23.6%) donkeys with titers of 1:25 in 66, 1:50 in 54, 1:100 in 49, and 1:200 in 5, and seroprevalence varied in 13 different regions, ranging from 9.1% to 32.6%.ConclusionsThe results of the present study indicated that the rate of infection with T. gondii in horses and donkeys is a little high in Liaoning province, northeastern China in comparison to other surveys in China, which suggests that consumption of horse and donkey meat in this area may represent a potential source for human infection with T. gondii.

Isolation and genetic characterization of H3N8 equine influenza virus from donkeys in China

During the 2007 outbreak of equine influenza (EIV) in China, an influenza virus designated A/donkey/Xinjiang/5/2007 (donkey/Xinjiang/2007) was isolated from a symptomatic donkey in Xinjiang Uygur Autonomous Region, China. To analyze the genetic evolution of the new isolate, the hemagglutinin (HA) gene of donkey/Xinjiang/2007 was amplified and sequenced. Sequence alignment, prediction of glycosylation sites and phylogenetic analysis of the HA1 protein of donkey/Xinjiang/2007 showed most similarity to the Florida sublineage clade 2 of the American lineage of equine influenza viruses. The HA1 sequence of donkey/Xinjiang/2007 had high identity with the Chinese EIV strains isolated in recent years and had low identity with A/swine/Anhui/1/2006, a horse-derived influenza virus isolated from pigs in China. Compared with vaccine strains, the HA1 amino acid sequence of donkey/Xinjiang/2007 varied in the five putative antigenic sites to different degrees and distributed in a different branch of the HA1 phylogenetic tree. Our findings provide further evidence of an American H3N8 equine influenza virus in China in addition to the European H3N8 equine influenza virus. Accordingly, a vaccine against equine influenza in China should contain components of both strains of viruses to prevent widespread infection.

African Maternal Origin and Genetic Diversity of Chinese Domestic Donkeys

The origin of domestic donkeys in China has been controversial. To clarify the origin of Chinese domestic donkeys, we investigated the partial mitochondrial D-loop sequences of 126 samples from 12 native breeds. The results revealed two mitochondrial origins, lineage Somali and lineage Nubian of African wild ass detected in Chinese domestic donkeys. Lineage Somali was predominant in Chinese domestic donkey breeds. The pattern of genetic variation in ass mtDNA D-loop sequences indicated that the two lineages Somali and Nubian from China had undergone population expansion events. In a combined analysis of lineages Somali and Nubian between previously published sequences from other countries/regions and sequences of Chinese domestic donkeys, the results indicated that the two lineages of Chinese domestic donkeys were from Africa and supported the African maternal origins of Chinese domestic donkeys. There was no obvious geographical structure in Chinese domestic donkey breeds, but the population showed abundant mtDNA diversity. The spread routes of Chinese domestic donkeys were also discussed.