Zoonotic Enteroparasites of Macaca Fascicularis In Palawan, Philippines

The expansion of ecotourism and forest encroachment in the Philippines creates a high-risk interface where human-macaque interactions occur at rates were cross-species transmission of disease may occur more frequently than previously known. Puerto Princesa Subterranean River National Park is a prime tourist destination in the country where long-tailed macaques live as commensals to humans. This study was conducted to assess zoonotic enteroparasites of Macaca fascicularis to determine their prevalence in the extant population. Fecal samples were collected during two-kilometer transect walks whilst opportunistic sampling was also conducted in the park proper where there is high tourist traffic. Among protozoans, Entamoeba coli showed the highest prevalence (34.29%), followed by Entamoeba spp. and Iodamoeba butschlii (31.43%), Endolimax nana (28.57%), Blastocystis sp. (22.86%), Chilomastix mesnili Entamoeba polecki (20%), and Giardia intestinalis (8.57%). From the helminth group, hookworm larva was the most prevalent (40%), followed by hookworm/strongylids ova (34.29%), Strongyloides sp. larva (28.57%), T. trichiura (20%), Ascaris sp. (11.43%), and lastly, Hymenolepis nana and Enterobius vermicularis (2.86%). This study demonstrates the importance of long-tailed macaques in the transmission of enteroparasites in an environment where there is frequent contact between nonhuman primates and people.

Parasitic Intestinal Protists of Zoonotic Relevance Detected in Pigs by Metabarcoding and Real-Time PCR

Several parasite species are shared between humans and pigs. We explored the application of next-generation sequencing-based metabarcoding supplemented with real-time PCR to fecal DNAs from 259 samples from 116 pigs in Denmark to detect and differentiate single-celled intestinal parasites of zoonotic relevance. Enterocytozoon bieneusi, Balantioides coli, and Giardia duodenalis were observed in 34/37 (92%), 148/259 (57%), and 86/259 (33%) samples, respectively. Entamoeba polecki ST1, E. polecki ST3, and Entamoeba hartmanni were detected in 104/259 (40%), 161/259 (62%), and 8/259 (3%) samples, respectively. Metabarcoding and real-time PCR detected Cryptosporidium in 90/259 (35%) and 239/259 (92%) of the samples, respectively, with Cryptosporidium suis and Cryptosporidium scrofarum observed in nearly equal proportions. Blastocystis subtypes 1, 3, 5, and 15 were found in 72 (28%), 6 (2%), 176 (68%), and 36 (14%) of 259 samples, respectively. Iodamoeba was identified in 1/259 samples (<1%), while none of 37 tested samples was positive for Dientamoeba fragilis. Our results illustrate how metabarcoding exemplifies a ‘one-fits-many’ approach to detecting intestinal single-celled parasites in feces supplemented with real-time PCR for selected parasites. Using metabarcoding with pathogen-specific assays may help detect emerging and previously underdetected pathogens and further elucidate the role of micro-eukaryotic parasites in human and animal health and disease.

Gastrointestinal parasites among swine bred in Edo State, Nigeria

Background: Swine production in Nigeria is encountering several constraints among which are diseases. This study was designed to determine the prevalence of gastrointestinal parasites of swine with respect to sex, age, location and hygiene practices in the breeding sites in three piggery farms; University of Benin Animal Farm; Osasio Farm Uselu in Egor Local Government Area (LGA), and Ojemai Farm Ekehuanwa Road, Oredo LGA, Benin City, Edo State, Nigeria. Methods: A total of 150 pigs were sampled from the 3 piggery farms. Faecal sample (10g each) was aseptically collected from the rectum of each selected pig and processed by concentration method to microscopically identify parasites in both saline and iodine preparations. A designed pre-tested structured questionnaire was interviewer-administered to each piggery owner to collect information on husbandry practices, animal health care issues and potential risk factors associated with parasitosis. Data was analysed using SPPSS version 20 software, while odd ratios (OR) with 95% confidence intervals (CI) were calculated on each potential risk factors. Results: Of the 150 pigs examined, 130 (86.6%) were infected with five gastrointestinal parasitic agents. Strongyloides ransomi 81 (54%) was the most frequently identified parasite, followed by Ascaris suum 68 (45.3%), Giardia lamblia 31 (20.6%), Entamoeba polecki 10 (6.6%) and Trichuris suis 10 (6.6%). Single and mixed infections were not significantly associated with the observed prevalence (p>0.05). Similarly, the sex of pig was not significantly associated with the prevalence of parasites (p=0.8824). The prevalence of parasitosis among the grower (87.8%) was not significantly different from the adult swine (87.5%) (p>0.05). Osasio had the highest infection rate (90.0%) but this rate was not significantly different from the two other farms (p>0.05). Conclusion: This study confirms high prevalence of gastrointestinal parasitic infections in pigs bred in Edo State, Nigeria. It is therefore recommended that farmers improve on their biosecurity and adhere to routine de-worming regimen of pigs. Keywords: age, gender, prevalence, gastrointestinal, parasite, swine French title: Parasites gastro-intestinaux chez les porcs élevés dans l'État d'Edo, au Nigéria Contexte: La production porcine au Nigéria rencontre plusieurs contraintes parmi lesquelles les maladies. Cette étude a été conçue pour déterminer la prévalence des parasites gastro-intestinaux des porcs en fonction du sexe, de l'âge, de l'emplacement et des pratiques d'hygiène dans les sites de reproduction de trois porcheries; Ferme animale de l'Université du Bénin; Osasio Farm Uselu dans la région du gouvernement local d'Egor (LGA) et Ojemai Farm Ekehuanwa Road, Oredo LGA, Benin City, État d'Edo, Nigéria. Méthodes: Au total, 150 porcs ont été échantillonnés dans les 3 fermes porcines. Un échantillon fécal (10 g chacun) a été prélevé de manière aseptique dans le rectum de chaque porc sélectionné et traité par une méthode de concentration pour identifier au microscope les parasites dans les préparations salines et d'iode. Un questionnaire structuré pré-testé conçu a été administré par un intervieweur à chaque propriétaire de porcherie pour recueillir des informations sur les pratiques d'élevage, les problèmes de santé animale et les facteurs de risque potentiels associés à la parasitose. Les données ont été analysées à l'aide du logiciel SPPSS version 20, tandis que les rapports impairs (OR) avec des intervalles de confiance (IC) à 95% ont été calculés sur chacun des facteurs de risque potentiels. Résultats: Sur les 150 porcs examinés, 130 (86,6%) ont été infectés par cinq agents parasitaires gastro-intestinaux. Strongyloides ransomi 81 (54%) était le parasite le plus fréquemment identifié, suivi par Ascaris suum 68 (45,3%), Giardia lamblia 31 (20,6%), Entamoeba polecki 10 (6,6%) et Trichuris suis 10 (6,6%). Les infections simples et mixtes n'étaient pas associées de manière significative à la prévalence observée (p>0,05). De même, le sexe du porc n'était pas significativement associé à la prévalence des parasites (p=0,8824). La prévalence de la parasitose chez le producteur (87,8%) n'était pas significativement différente de celle des porcs adultes (87,5%) (p>0,05). Osasio avait le taux d'infection le plus élevé (90,0%) mais ce taux n'était pas significativement différent des deux autres fermes (p>0,05). Conclusion: Cette étude confirme la forte prévalence des infections parasitaires gastro-intestinales chez les porcs élevés dans l'État d'Edo, au Nigeria. Il est donc recommandé que les éleveurs améliorent leur biosécurité et adhèrent au régime vermifuge de routine des porcs. Mots-clés: âge, sexe, prévalence, gastro-intestinal, parasite, porcin

Prevalence and Genetic Identification of Three Entamoeba Species in Pigs in Southeastern China

Parasitic Entamoeba spp. can infect many classes of vertebrates including humans and pigs. Entamoeba suis and zoonotic Entamoeba polecki have been identified in pigs, and swine are implicated as potential reservoirs for Entamoeba histolytica. However, the prevalence of Entamoeba spp. in pigs in southeastern China has not been reported. In this study, 668 fecal samples collected from 6 different regions in Fujian Province, southeastern China, were analyzed to identify three Entamoeba species by nested PCR and sequencing analysis. The overall prevalence of Entamoeba spp. was 55.4% (370/668; 95% CI 51.6% to 59.2%), and the infection rate of E. polecki ST1 was the highest (302/668; 45.2%, 95% CI 41.4% to 49.0%), followed by E. polecki ST3 (228/668; 34.1%, 95% CI 30.5% to 37.7%) and E. suis (87/668; 13.0%, 95% CI 10.5% to 15.6%). E. histolytica was not detected in any samples. Moreover, the coinfection rate of E. polecki ST1 and ST3 was 25.1% (168/668; 95% CI 21.9% to 28.4%), the coinfection rate of E. polecki ST1 and E. suis was 3.7% (25/668; 95% CI 2.3% to 5.2%), the coinfection rate of E. polecki ST3 and E. suis was 0.3% (2/668), and the coinfection rate of E. polecki ST1, E. polecki ST3, and E. suis was 4.0% (27/668; 95% CI 2.5% to 5.5%). A representative sequence (MK347346) was identical to the sequence of E. suis (DQ286372). Two subtype-specific sequences (MK357717 and MK347347) were almost identical to the sequences of E. polecki ST1 (FR686383) and ST3 (AJ566411), respectively. This is the first study to survey the occurrence and to conduct molecular identification of three Entamoeba species in southeastern China. This is the first report regarding mixed infections with E. suis, E. polecki ST1, and E. polecki ST3 in China. More research studies are needed to better understand the transmission and zoonotic potential of Entamoeba spp.

Coinfection with Entamoeba polecki and Brachyspira hyodysenteriae in a pig with severe diarrhea

Enteric disease in pigs is usually of multifactorial etiology, including infectious and non-infectious factors. In many cases of endemic diarrhea in weaner-to-finisher pigs, the combination of 2 or more microorganisms leads to aggravation of intestinal lesions and, consequently, clinical signs. We autopsied a 4-mo-old fattening pig with diarrhea and diagnosed severe fibrinonecrotizing typhlocolitis. Numerous spiral-shaped bacteria and amoeba-like PAS-positive protozoa were observed in the cecal and colonic mucosa and submucosa. Brachyspira hyodysenteriae was detected by PCR from colonic content. By in situ hybridization, large numbers of Entamoeba polecki were found within the lamina propria and submucosa; moderate numbers of Blastocystis sp. and scattered trichomonads were present in intestinal content. In addition, Entamoeba polecki, Balantidium spp., Blastocystis sp., and Trichomonas sp. were also detected by PCR.

Molecular identification ofEntamoebaspecies in savanna woodland chimpanzees (Pan troglodytes schweinfurthii)

SUMMARYTo address the molecular diversity and occurrence of pathogenic species of the genusEntamoebaspp. in wild non-human primates (NHP) we conducted molecular-phylogenetic analyses onEntamoebafrom wild chimpanzees living in the Issa Valley, Tanzania. We compared the sensitivity of molecular [using a genus-specific polymerase chain reaction (PCR)] and coproscopic detection (merthiolate-iodine-formaldehyde concentration) ofEntamoebaspp. We identifiedEntamoebaspp. in 72 chimpanzee fecal samples (79%) subjected to species-specific PCRs for sixEntamoebaspecies/groups (Entamoeba histolytica, Entamoeba nuttalli, Entamoeba dispar, Entamoeba moshkovskii, Entamoeba coliandEntamoeba poleckiST2). We recorded threeEntamoebaspecies:E. coli(47%),E. dispar(16%),Entamoeba hartmanni(51%). Coproscopically, we could only distinguish the cysts of complexE. histolytica/dispar/moshkovskii/nuttalliandE. coli. Molecular prevalence of entamoebas was higher than the prevalence based on the coproscopic examination. Our molecular phylogenies showed that sequences ofE. disparandE. colifrom Issa chimpanzees are closely related to sequences from humans and other NHP from GenBank. The results showed that wild chimpanzees harbourEntamoebaspecies similar to those occurring in humans; however, no pathogenic species were detected. Molecular-phylogenetic methods are critical to improve diagnostics of entamoebas in wild NHP and for determining an accurate prevalence ofEntamoebaspecies.