Predominance of the Blastocystis subtype ST5 among free-living sympatric rodents within pig farms in China suggests a novel transmission route from farms.

Blastocystis is a parasitic protist that can infect humans and various domestic and wild animals. However, there is limited research on the prevalence of this parasite among rodents, particularly those living in pig farm settings. Therefore, to investigate the occurrence, molecular characterization, and zoonotic potential of Blastocystis among rodents within pig farm environments, we conducted an investigation of 227 rodents and shrews from 34 pig farms located in Henan, Shaanxi, and Shanxi provinces of China using nested PCR of the SSU rRNA gene of Blastocystis. The potential transmission and public health implications were also assessed from a One Health perspective. Blastocystis was detected in 86 (37.9%) fecal samples. The highest infection rate was observed among Ruttus norvegicus (73.7%, 42/58), followed by Ruttus tanezumi (30.1%, 41/136), and Mus musculus (12.0%, 3/25). However, it was not detected among individuals with Apodemus agrarius (n = 1) and Crocidura shantungensis (n = 7). Five known zoonotic Blastocystis subtypes (ST1-ST5) were identified, with ST4 (51.2%, 44/86) and ST5 (40.7%, 35/86) being the predominant ones, followed by ST1 (3.5%, 3/86), ST3 (3.5%, 3/86), and ST2 (1.2%, 1/86). ST4 was prevalent among R. norvegicus (83.3%, 35/42), while ST5 dominated R. tanezumi (70.7%, 29/41). Furthermore, ST5 exhibited the widest distribution at pig farm level, accounting for 65.0% (13/20) of Blastocystis-positive pig farms. This investigation presents the first documented Blastocystis infection in R. tanezumi and M. musculus, highlighting the predominant presence of the zoonotic ST5 subtype in rodents for the first time. The results demonstrate that sympatric rodents can serve as natural reservoirs for Blastocystis and play a role in its transmission. These findings provide information on the dynamics of rodent transmission and emphasize the potential public health threat posed by zoonotic Blastocystis subtypes spillover from pig farms.

Morphological and molecular biology identification of Cystoisospora sp. in the blue fox, Alopex lagopus (Linnaeus, 1758).

To investigate the prevalence and molecular characteristics of Cystoisospora sp. in blue fox (Alopex lagopus), Sheather's sugar floatation method was conducted to detect coccidia in 423 fresh fecal samples randomly collected from blue fox farms from three cities in China. The overall prevalence of coccidia was 1.4% (6/423), and three Cystoisospora sp. (Cystoisospora fennechi, Cystoisospora sp. I and Cystoisospora vulpina) were identified by their morphological characteristics. The 18S ribosomal RNA (rRNA) and cytochrome c oxidase subunit I (COI) locus sequences were sequenced for molecular biological identification, homology comparison, and phylogenetic analysis of Cystoisospora sp. by single-oocyst selection technology and multi-locus-nested PCR amplification. At the 18S rRNA and COI loci, C. vulpina had 99.48% and 99.59% homology, respectively, with Cystoisospora canis and Cystoisospora ohioensis from canines. Phylogenetic analysis indicated that C. vulpina was clustered in a clade with Cystoisospora sp. from Canidae, which the relatives are consistent with the hosts. To our knowledge, this is the first report on molecular identification and evolutionary analysis of C. vulpina at two different loci.

Acaricidal and repellent activities of ethanol extracts of nine chinese medicinal herbs against Rhipicephalus microplus (Acari: Ixodidae).

Rhipicephalus microplus is a major threat to the cattle industry worldwide. The intensive use of acaricides and repellents has resulted in drug resistance. Hence, effective and eco-friendly pest control alternatives are urgently needed, especially from natural plant resources. In this study, the acaricidal and repellent activities of nine herbs against the larvae and eggs of R. microplus were evaluated. The results showed that ethanol extracts of star anise (Illicium verum), chaulmoogra (Hydnocarpus anthelmintica), motherwart (Leonurus artemisia), mandarin orange peel (citri reticulatae pericarpium, i.e., peel of Citrus reticulata fruit), and stemona (Stemona sessilifolia) had good contact acaricidal activities of 100, 98, 94, 88 and 86%, respectively, whereas star anise and clove (Syzygium aromaticum) had good fumigant acaricidal activities of 98 and 96%, respectively. The hatching inhibition rate of star anise against R. microplus eggs was 100%. All nine herbs had good real-time repellent rates, but only castor bean and star anise had repellent effects after 48 h (81.3 and 79.6%, respectively). This is the first report of the acaricidal and repellent activities of these medicinal herbs against R. microplus. Ethanol extracts of these herbs might be considered as potential alternatives to chemical acaricides for control of R. microplus.

Morphological and molecular characteristics of a single oocyst for the identification of Eimeria species in domestic rabbits (Oryctolagus cuniculus f. domesticus).

Coccidiosis caused by Eimeria is one of the most common diseases in domestic rabbits (Oryctolagus cuniculus f. domesticus), with 11 Eimeria species in domestic rabbits recognized internationally. To identify Eimeria species more accurately, a method based on the molecular characteristics of a single oocyst with multiple gene loci was established by combining morphological and molecular biology. The results showed that the total infection rate of Eimeria in domestic rabbits was 44.2 % (152/344). Ten Eimeria species were identified in domestic rabbits based on morphological characteristics, namely Eimeria vejdovskyi (39.5 %, 136/344), E. magna (18.0 %, 62/344), E. perforans (17.4 %, 60/344), E. intestinalis (12.5 %, 43/344), E. media (11.9 %, 41/344), E. coecicola (4.4 %, 15/344), E. irresidua (3.8 %, 13/344), E. exigua (2.6 %, 9/344), E. stiedai (2.3 %, 8/344), and E. piriformis (1.5 %, 5/344). The molecular biological identification of Eimeria in domestic rabbits was conducted through single oocyst selection and nested polymerase chain reaction amplification with multiple gene loci. We obtained the sequences of the 18S rRNA, ITS-1 and COI gene loci of E. magna, E. perforans, E. vejdovskyi, E. media, E. intestinalis, and E. coecicola. The results showed that the molecular biology and morphological identification results of single oocysts were consistent and could be used for the molecular identification of Eimeria at the single oocyst level. This study provides an efficient tool for identification of Eimeria in domestic rabbits and the population genetic study of Eimeria in domestic rabbits.

The global prevalence of parasites in non-biting flies as vectors: a systematic review and meta-analysis.

BACKGROUND: Non-biting flies such as the house fly (Musca domestica), the Australian sheep blowfly (Lucilia cuprina) and the oriental latrine fly (Chrysomya megacephala) may carry many parasites. In the present study, we performed a systematic overview of the different species of parasites carried by non-biting flies, as well as of isolation methods, different geographical distribution, seasonality and risk assessment. METHODS: A meta-analysis was carried out with the aim to review the global prevalence of parasite transmission in non-biting flies. A total sample size of 28,718 non-biting flies reported in studies worldwide satisfied the predetermined selection criteria and was included in the quantitative analysis. RESULTS: The global prevalence of parasites in non-biting flies was 42.5% (95% confidence interval [CI] 31.9-53.2%; n = 15,888/28,718), with the highest prevalence found for non-biting flies in Africa (58.3%; 95% CI 47.4-69.3%; n = 9144/13,366). A total of 43% (95% CI 32.1-54.4%; n = 7234/15,282) of house flies (M. domestica), the fly species considered to be the most closely associated with humans and animals, were found with parasites. The prevalence of parasites in the intestine of non-biting flies was 37.1% (95% CI 22.7-51.5%; n = 1045/3817), which was significantly higher than the prevalence of parasites isolated from the body surface (35.1%; 95% CI 20.8-49.4%; n = 1199/3649; P < 0.01). Of the 27 reported parasites, a total of 20 known zoonotic parasites were identified, with an infection rate of 38.1% (95% CI 28.2-48.0%; n = 13,572/28,494). CONCLUSIONS: This study provides a theoretical basis for the public health and ecological significance of parasites transmitted by non-biting flies.

Molecular characterization of Cryptosporidium spp., Enterocytozoon bieneusi and Giardia duodenalis in laboratory rodents in China.

Cryptosporidium spp., Enterocytozoon bieneusi and Giardia duodenalis are significant zoonotic intestinal pathogens that can cause gastrointestinal symptoms such as diarrhea and induce a host immune response. A total of 1237 fecal samples were collected from laboratory rodents (rats, mice and guinea pigs) from four different locations in China to investigate the infection rates and molecular characterization of these pathogens on experimental animals. Genomic DNA was extracted from each sample, and PCR amplifications were done. Overall, the Cryptosporidium spp. infection rate was 3.8% (47/1237). Four known Cryptosporidium species were identified, namely C. parvum, C. muris, C. tyzzeri and C. homai, the three former being zoonotic species. The overall E. bieneusi infection rate was 3.0% (37/1237). Seven known E. bieneusi genotypes, namely S7, BEB6, J, Henan-IV, CHG10, D and WL6, were detected by sequence analysis. Among these, genotypes D, Henan-IV and CHG10 have a high zoonotic risk. Giardia duodenalis was not detected at any of the three loci (SSU rRNA, bg and gdh) after PCR amplification. This study provides basic data for these pathogens in laboratory rodents in China and lays the foundation for their prevention and control in laboratory animals.

Title: Caractérisation moléculaire de Cryptosporidium spp., Enterocytozoon bieneusi et Giardia duodenalis chez des rongeurs de laboratoire en Chine. Abstract: Cryptosporidium spp., Enterocytozoon bieneusi et Giardia duodenalis sont des agents pathogènes intestinaux zoonotiques importants, qui peuvent provoquer des symptômes gastro-intestinaux tels que la diarrhée et induire une réponse immunitaire de l'hôte. Au total, 1237 échantillons fécaux ont été prélevés sur des rongeurs de laboratoire (rats, souris et cobayes) de quatre endroits différents en Chine pour étudier les taux d'infection et la caractérisation moléculaire de ces agents pathogènes sur des animaux de laboratoire. L'ADN génomique a été extrait de chaque échantillon et des amplifications par PCR ont été effectuées. Dans l'ensemble, le taux d'infection par Cryptosporidium spp. était de 3,8 % (47/1237). Quatre espèces connues de Cryptosporidium ont été identifiées, à savoir C. parvum, C. muris, C. tyzzeri et C. homai, les trois premières étant des espèces zoonotiques. Le taux global d'infection par E. bieneusi était de 3,0 % (37/1 237). Sept génotypes connus d'E. bieneusi, à savoir S7, BEB6, J, Henan-IV, CHG10, D et WL6, ont été détectés par analyse de séquence. Parmi ceux-ci, les génotypes D, Henan-IV et CHG10 ont un risque zoonotique élevé. Giardia duodenalis n'a été détecté à aucun des trois loci (SSU ARNr, bg et gdh) après amplification par PCR. Cette étude fournit des données de base sur ces agents pathogènes chez les rongeurs de laboratoire en Chine et pose les bases de leur prévention et de leur contrôle chez les animaux de laboratoire.