Molecular characterization and zoonotic potential of Entamoeba spp., Enterocytozoon bieneusi and Blastocystis from captive wild animals in northwest China.

BACKGROUND: Parasites Entamoeba spp., Enterocytozoon bieneusi and Blastocystis are prevalent pathogens causing gastrointestinal illnesses in animals and humans. Consequently, researches on their occurrence, distribution and hosts are crucial for the well-being of both animals and humans. Due to the confined spaces and frequent interaction between animals and humans, animal sanctuaries have emerged as potential reservoirs for these parasites. In this study, the wildlife sanctuary near the Huang Gorge of the Qinling Mountains in northwest China is chosen as an ideal site for parasite distribution research, considering its expansive stocking area and high biodiversity. RESULTS: We collected 191 fecal specimens from 37 distinct wildlife species and extracted genomic DNA. We identified these three parasites by amplifying specific gene regions and analyzed their characteristics and evolutionary relationships. All the parasites exhibited a high overall infection rate, reaching 90.05%. Among them, seven Entamoeba species were identified, accounting for a prevalence of 54.97%, with the highest infection observed in Entamoeba bovis. In total, 11 Enterocytozoon bieneusi genotypes were discovered, representing a prevalence of 35.08%, including three genotypes of human-pathogenic Group 1 and two novel genotypes (SXWZ and SXLG). Additionally, 13 Blastocystis subtypes were detected, showing a prevalence of 74.87% and encompassing eight zoonotic subtypes. All of the above suggests significant possibilities of parasite transmission between animals and humans. CONCLUSIONS: This study investigated the occurrence and prevalence of three intestinal parasites, enhancing our understanding of their genetic diversity and host ranges in northwest China. Furthermore, the distribution of these parasites implies significant potential of zoonotic transmission, underscoring the imperative for ongoing surveillance and implementation of control measures. These efforts are essential to mitigate the risk of zoonotic disease outbreaks originating from wildlife sanctuary.

Epidemiological investigation of Entamoeba in wild rhesus macaques in China: A novel ribosomal lineage and genetic differentiation of Entamoeba nuttalli.

Wild rhesus macaques are a potential source of zoonotic parasites for humans, and Entamoeba spp. are common intestinal parasites. To investigate the prevalence of Entamoeba in wild rhesus macaques in China and explore the genetic differentiation of the potentially pathogenic species Entamoeba nuttalli, a total of 276 fecal samples from five populations at high altitudes (HAG, 2,800-4,100 m above sea level) and four populations at low altitudes (LAG, 5-1,000 m above sea level) were collected. PCR methods based on the ssrRNA gene were used to detect Entamoeba infection. Genotyping of E. nuttalli was performed based on six tRNA-linked short tandem repeat (STR) loci for further genetic analyses. The results revealed that Entamoeba infection (69.2%) was common in wild rhesus macaques in China, especially in LAG which had a significantly higher prevalence rate than that in HAG (P < 0.001). Three zoonotic species were identified: Entamoeba chattoni (60.9%) was the most prevalent species and distributed in all the populations, followed by Entamoeba coli (33.3%) and Entamoeba nuttalli (17.4%). In addition, a novel Entamoeba ribosomal lineage named RL13 (22.8%) was identified, and phylogenetic analysis revealed a close genetic relationship between RL13 and Entamoeba. hartmanni. Genotyping of E. nuttalli obtained 24 genotypes from five populations and further analysis showed E. nuttalli had a high degree of genetic differentiation (FST > 0.25, Nm < 1) between the host populations. The result of analysis of molecular variance (AMOVA) revealed that observed genetic differences mainly originate from differences among populations (FST = 0.91). Meanwhile, the phylogenetic tree showed that these genotypes of E. nuttalli were clustered according to geographical populations, indicating a significant phylogeographic distribution pattern. Considering the potential pathogenicity of E. nuttalli, attention should be paid to its risk of zoonotic transmission.

Molecular evidence suggests the occurrence of Entamoeba moshkovskii in pigs with zoonotic potential from eastern India.

Entamoeba moshkovskii Tshalaia, 1941 is prevalent in developing countries and it is considered to be primarily a free-living amoeba, which is morphologically indistinguishable, but biochemically and genetically different from the human infecting, pathogenic Entamoeba histolytica Schaudinn, 1903. The pathogenic potential of this organism is still under discussion. Entamoeba moshkovskii in human stool samples has been reported in different countries such as the United States, Italy, Australia, Iran, Turkey, Bangladesh, India (Pondicherry), Indonesia, Colombia, Malaysia, Tunisia, Tanzania and Brazil, but no data are available about the occurrence of E. moshkovskii in farm animals. This study provides data on the occurrence of E. moshkovskii in pigs in a total of 294 fresh faecal samples collected from five different regions in Kolkata, West Bengal, India. Stool samples were tested by nested PCR using primers targeting SSU rDNA of E. moshkovskii. The amplified PCR products were further confirmed by RFLP technique. Purified nested PCR products were also sequenced and identified via BLAST program run on the NCBI website to confirm species along with their genetic characteristics of the E. moshkovskii isolates. Overall 5.4 % samples were identified as E. moshkovskii positive. Results of this study demonstrate that swine can host E. moshkovskii and should be considered as a potential natural reservoir for E. moshkovskii. However, the occurrence of E. moshkovskii infection in pigs was not statistically associated with their faecal consistency, sex and developmental stage.

Management of Entamoeba histolytica in the non-human primates at the Singapore Zoo.

Amebic dysentery caused by Entamoeba histolytica accounts for significant morbidity in the non-human primates (NHP) at the Singapore Zoo. This includes the animals in the collection as well as a sizeable free-roaming wild crab-eating macaque (Macaca fascicularis) population. The disease is of great concern because of its zoonotic potential. Passive surveillance, both ante and post-mortem, of NHP displaying clinical symptoms and active surveillance of NHP assessed to be at a higher risk of infection were carried out via fecal real-time polymerase chain reaction (PCR) testing for 4 years. Treatment of the disease with 25 mg/kg metronidazole BID for 10 days followed by 15 mg/kg paromomycin BID for 7 days achieved good clinical resolution in most cases that tested positive. Three diseased NHP with severe clinical signs of weight loss, lethargy, and diarrhea were anesthetized for veterinary diagnostic investigation. Mesenteric lymphadenopathy was consistently seen on ultrasound examination in these severe cases of entamoebiasis. Two animals eventually died of severe chronic enteritis due to the disease. The eradication of entamoebiasis in the NHP at the Singapore Zoo may be complicated by the maintenance of a disease reservoir in wildlife, but a combination of timely treatment and efforts at maintaining biosecurity can help manage the disease in the collection.

Diversity, geographical distribution, and prevalence of Entamoeba spp. in Brazil: a systematic review and meta-analysis.

The genus Entamoeba includes a variety of widely distributed species adapted to live in the digestive tracts of humans and a large variety of animals of different classes. The objective of this study was to investigate the prevalence, distribution, and molecular epidemiology of Entamoeba spp. in different classes of hosts in Brazil. Studies that analyzed hosts from several classes, including humans and domestic, wild, or captive animals, were considered. The pooled prevalence of Entamoeba spp. was calculated using the random-effects model. A total of 166 studies on humans and 16 on animals were included. The prevalence of Entamoeba spp. in the Brazilian population was 22% (95% CI: 21-24). The state with the highest prevalence was Paraiba with 72%, followed by Federal District with 53%, and Rondonia with 50%. In immunocompromized patients, the prevalence was 18%, and cancer (36%) was the most prevalent cause of immunosuppression. The prevalence of Entamoeba spp. in animal hosts was 12% (95% CI: 7-17). Captive wild animals and domestic farm animals showed the highest prevalence, with 16% and 15%, respectively. The species found more often were E. coli (86.5%), E. dispar (7.9%), and E. histolytica (3.1%). In conclusion, a high prevalence (22%) of Entamoeba spp. was found in the Brazilian population, with a prevalence of up to 50% mainly in the northern, northeastern, and central-western regions. The pathogenic species E. histolytica is distributed in most Brazilian regions, with significant prevalence percentages. Among animals, unidentified Entamoeba species were most prevalent in mammals.

TITLE: Diversité, répartition géographique et prévalence d'Entamoeba spp. au Brésil : revue systématique et méta-analyse. ABSTRACT: Le genre Entamoeba comprend une variété d'espèces largement distribuées, adaptées à vivre dans le tube digestif des humains et une grande variété d'animaux de différentes classes. L'objectif de cette étude était d'étudier la prévalence, la distribution et l'épidémiologie moléculaire d'Entamoeba spp. dans différentes classes d'hôtes au Brésil. Les études qui ont analysé les hôtes de plusieurs classes, y compris les humains et les animaux domestiques, sauvages ou captifs, ont été prises en compte. La prévalence combinée d'Entamoeba spp. a été calculée à l'aide du modèle à effets aléatoires. Au total, 166 études sur l'homme et 16 sur les animaux ont été incluses. La prévalence d'Entamoeba spp. dans la population brésilienne était de 22 % (IC à 95 % : 21­24). L'état avec la prévalence la plus élevée était Paraiba avec 72 %, suivi du District fédéral avec 53 % et Rondonia avec 50 %. Chez les patients immunodéprimés, la prévalence était de 18 % et le cancer (36 %) était la cause la plus fréquente d'immunosuppression. La prévalence d'Entamoeba spp. chez les hôtes animaux était de 12 % (IC à 95 % : 7­17). Les animaux sauvages en captivité et les animaux domestiques d'élevage ont affiché la prévalence la plus élevée, avec respectivement 16 % et 15 %. Les espèces trouvées le plus souvent étaient E. coli (86,5 %), E. dispar (7,9 %) et E. histolytica (3,1 %). En conclusion, une prévalence élevée (22 %) d'Entamoeba spp. a été trouvée dans la population brésilienne, allant jusqu'à 50 % principalement dans les régions du nord, du nord-est et du centre-ouest. L'espèce pathogène E. histolytica est répartie dans la plupart des régions du Brésil, avec des pourcentages de prévalence importants. Parmi les animaux, les espèces d'Entamoeba non identifiées étaient les plus répandues chez les mammifères.

First report of Eimeria and Entamoeba infection in alpacas (Vicugna pacos) in Shanxi Province, northern China.

Intestinal protozoa Eimeria and Entamoeba can infect many animal species including alpacas. However, data on the prevalence and pathogenicity of species of the two genera Eimeria and Entamoeba in alpacas in China is scarce. The current study was carried out to investigate the prevalence of Eimeria and Entamoeba in alpacas in two cities (Taiyuan and Xinzhou) in Shanxi Province, northern China, using PCR-based approaches. Eimeria spp. were only found in Taiyuan city, and the overall prevalence was 1.64%. All samples collected from male alpacas were PCR-negative for Eimeria. Four Eimeria-positive samples were tested positive as Eimeria lamae. The molecular prevalence of Entamoeba in alpacas was 18.03% (66/366), including 16.39% (50/305) in alpacas from Taiyuan city and 26.23% (16/61) from Xinzhou city, respectively. The Entamoeba prevalence in male alpacas (25.00%) was significantly higher than that in female alpacas (15.69%). Entamoeba bovis was the predominant species, and no Entamoeba histolytica infection was detected. Nine unique SSU rRNA gene sequences of Entamoeba were obtained which formed a new cluster. The results showed that sex and location might be the risk factors associated with prevalence of Eimeria spp., and sex might be the risk factor associated with prevalence of Entamoeba spp.. This is the first report of Entamoeba in alpacas worldwide. These findings expand our understanding of the prevalence and genetic diversity of Eimeria and Entamoeba in alpacas.

Diversity of parasites in two captive chimpanzee populations in southern Gabon.

Captive chimpanzees living in confined environments like sanctuaries or primatology centers are frequently affected by gastrointestinal parasites. Some of these are likely to be transmitted to humans and may seriously affect public health. However little information is currently available on the gastrointestinal parasites of primates living in such environments. Here, we characterize the diversity and prevalence of gastrointestinal parasites in two populations of captive chimpanzees living in south-eastern Gabon. Our study reveals that at least nine parasite species infect the chimpanzees with high prevalence, including several helminths (Ascaris spp., Enterobius spp., Strongyloides spp., Trichuris spp., Hymenolepis spp., Mammomonogamus spp), three protozoa (Balantioides spp., Entamoeba spp. and Troglodytella spp) and several unidentified parasites. All the parasite taxa we identified had previously been identified in other primates, including humans. Age, sex and site type may influence infection rates and/or parasite diversity found in a particular host.

Gastrointestinal entamoebiasis in captive anurans in North America.

Infections with Entamoeba spp. are recognized as a cause of clinical disease in many species including humans and reptiles; however, cases in amphibians are under-reported. Investigation of a mortality event among a captive population of Cranwell's horned frogs Ceratophrys cranwelli at a production facility in Florida, USA, revealed that deaths were due to the newly described Entamoeba species CT1. Infection caused severe necroulcerative gastroenterocolitis with a predilection for the colon. To date, this Entamoeba species has only been described in invasive cane toads Rhinella marina in Australia. Retrospective screening of archived anuran cases from a zoological pathology service identified 8 cases from captive populations that had histological evidence of gastrointestinal entamoebiasis. Molecular characterization was positive in 3 cases. Two cases, 1 in a Puerto Rican crested toad Peltophryne lemur and 1 in an Amazon milk frog Trachycephalus resinifictrix, showed 100% homology to E. ranarum and 1 case in a White's tree frog Litoria caerulea showed 100% homology to Entamoeba sp. CT1. This is the first report of novel Entamoeba sp. CT1 being associated with clinical disease in anurans within North America and also the first report of this Entamoeba species causing disease within managed collections as far back as 2003.

The first survey and molecular identification of Entamoeba spp. in farm animals on Qinghai-Tibetan Plateau of China.

Protozoans of Entamoeba spp. are globally distributed zoonotic parasites that infect diverse animal hosts and humans. Prevalence and species/genotypes distribution of Entamoeba spp. in domestic animals are not fully investigated on Qinghai-Tibetan Plateau (QTP), an animal husbandry and agriculture region of China. In a survey, 528 fecal samples were collected from 7 species of domestic animals on multiple locations across QTP region and analyzed by PCR and sequencing analysis. The overall prevalence of Entamoeba spp. infection in all examined animals was 97.9 %. Four Entamoeba species, E. bovis, E. moshkovskii, E. ecuadoriensis and E. histolytica were found, and majority (94.2 %) of Entamoeba-infected animals harbored E. bovis. Six Entamoeba genotypes, Entamoeba ribosomal lineages (RL) 1, 2, 3, 4, 8 and 9 were identified by sequencing analysis. Two zoonotic species, E. moshkovskii and E. histolytica, were present in horses, while E. ecuadoriensis and E. bovis were found in horses and all species of seven farm animals, respectively. It was also observed that six Entamoeba genotypes were distributed in animals in specific pattern. The results revealed high prevalence of Entamoeba spp. infection in livestock, broad range of hosts as well as diversity and species/genotype distribution of Entamoeba spp. in farm animals inhabiting on the high altitude QTP region.

Entamoeba ranarum Infection in a Ball Python (Python regius).

The pathogenic Entamoeba species in snakes is widely regarded to be Entamoeba invadens, which can cause severe amoebiasis with up to 100% mortality. In this case report, we describe a ball python (Python regius) that died after short-term weight loss. Necropsy revealed severe necrotizing colitis with large numbers of intralesional Entamoeba trophozoites. Molecular genetic analysis identified these trophozoites as Entamoeba ranarum, a parasite more usually found in amphibians. Furthermore, the extended history revealed that toads (Rhinella marina) had been housed together with the python. This report illustrates the danger of protozoal cross-infections in exotic animals as well as the importance of molecular genetic tools in Entamoeba diagnosis.

Occurrence and genetic identifications of porcine Entamoeba, E. suis and E. polecki, at Tangerang in West Java, Indonesia.

Entamoeba suis and E. polecki subtype (ST) 1 and ST3 recently have been inferred to be virulent in pigs. However, because relevant molecular epidemiological surveys have been limited, the prevalences of these species remain unknown and their pathogenicities are still controversial. We surveyed 196 fecal samples of pigs (118 of adults, 78 of piglets) at Tangerang in West Java, Indonesia, in 2017, employing PCR using porcine Entamoeba-specific primers. E. suis was the more frequently detected species, observed in 81.1% of samples, while E. polecki ST1 and ST3 were detected in 18.4% and 17.3% of samples, respectively; mixed infections (harboring 2-3 species or subtypes of Entamoeba) were confirmed in 29.3% of positive samples. Statistically significant differences in the positive rates were not seen between adult pigs and piglets, except for those of E. polecki ST3. The prevalences of Eimeria spp. and/or Cystoisospora suis (79.1%), strongyles (55.6%), and Strongyloides spp. (6.1%) were also observed morphologically in the samples. Further chronological or seasonal investigations of pigs and humans in these high-prevalence areas are needed to assess the virulence of the Entamoeba parasites, including the effects on pig productivity, and to evaluate the zoonotic impacts of these organisms.

Prevalence, molecular epidemiology and zoonotic risk of Entamoeba spp. from experimental macaques in Yunnan Province, southwestern China.

Amebiasis is a worldwide parasitic zoonosis, with symptoms of abdominal discomfort, indigestion, diarrhea, and even death. However, limited information about the prevalence of Entamoeba spp. in experimental nonhuman primates (NHPs) in southwestern China is available. The objective of the current study was to investigate the frequency and species identity of Entamoeba to evaluate potential zoonotic risk factors for Entamoeba spp. infection in experimental NHPs. A total of 505 fecal samples were collected from NHPs (macaques) and analyzed by PCR analysis the small subunit rRNA (SSU rRNA) gene of Entamoeba spp. Forty-seven specimens were positive for Entamoeba spp., and the prevalence of Entamoeba spp. was 9.31% (47/505). Significant differences in the prevalence rates among the three breeds (P = 0.002 < 0.01, df = 2, χ2 = 12.33) and feed types (P = 0.001 < 0.01, df = 1, χ2 = 10.12) were observed. Altogether, four Entamoeba species, including E. dispar (57.44%), E. chattoni (29.78%), E. histolytica (6.38%), and E. coli (6.38%), were identified by DNA sequence analysis. The results suggested a low prevalence but high diversity of Entamoeba species in experimental NHPs in Yunnan Province, southwestern China. Results of this study contribute to the knowledge of the genetic characteristics of Entamoeba spp. in NHPs.

Swine multifocal ulcerative colitis and crypt abscesses associated with Entamoeba polecki subtype 3 and Salmonella Typhimurium.

Piglets aged approximately 50 days exhibited diarrhea and wasting. Multiple white foci were detected in the colon of a dead piglet; histopathological findings revealed multifocal ulcers and crypt abscesses with Entamoeba trophozoites and gram-negative bacilli in the piglet. These pathogens were identified as Entamoeba polecki subtype 3 and Salmonella enterica serovar Typhimurium, respectively. Numerous E. polecki subtype 3 trophozoites were located on the edge of the ulcerative and necrotic lesions in the lamina propria. Crypt abscesses were associated with S. Typhimurium. These results suggest that E. polecki subtype 3 caused multifocal ulcerative colitis accompanied by crypt abscesses with S. Typhimurium in the piglet. This study is the first report of colitis with E. polecki subtype 3 and S. Typhimurium coinfection.

Copro-molecular study of Entamoeba infection among the indigenous community in Malaysia: A first report on the species-specific prevalence of Entamoeba in dogs.

BACKGROUND: Entamoeba is a free-living protozoan parasitic species that infect a variety of hosts. In humans, Entamoeba histolytica is the causative agent of amoebiasis. Entamoeba species has also been reported in dogs. However, little is known about the molecular epidemiology and the specific species of this parasite in dogs globally, including Malaysia. As dogs are important companion animals for the indigenous community, and close contact with dogs is part of the natural living conditions for this community, this study aims to determine the prevalence and molecular epidemiology of Entamoeba species in human and dogs in Malaysia. METHOD: The presence of Entamoeba species was examined in 504 fresh fecal samples, collected randomly from 411 humans and 93 dogs using microscopy and polymerase chain reaction (PCR) amplifying 16 s ribosomal RNA (rRNA). Data was analyzed using appropriate statistical analysis. RESULTS: The microscopy data showed an overall occurrence of Entamoeba species of 26.3% (108/411) and 36.6% (34/93) in humans and dogs respectively. In humans, the most common species was a single infection of E. dispar (26.5%; 13/49), followed by E. histolytica and E. moshkovskii, (20.4% for each species respectively). Double infection of E. dispar + E. moshkovskii was detected at 10.2%, followed by E. dispar + E. histolytica (8.2%) and E. moshkovskii and E. histolytica (6.1%). 8.2% of the samples had triple infection with all three species. In animals, E. moshkovskii (46.7%) was the most common species detected, followed by E. histolytica, and E. dispar, at 20.0% and 13.3% respectively. Double infection with E. moshkovskii + E. histolytica and a triple infection were found in 2 samples (13.3%) and 1 (6.7%) sample respectively. Risk factor analysis showed that members of the community who used untreated water were more prone to be infected with Entamoeba. CONCLUSION: This study provides information on the species-specific occurrence of Entamoeba infection, the potential risk factors and their zoonotic potential to humans. This is the first report to describe the molecular occurrence of Entamoeba species in dogs in Malaysia. The presence of pathogenic Entamoeba species implies that dogs could be a reservoir or mechanical host for human amoebiasis. Further studies need to be conducted to better understand the transmission dynamics and public health significance of Entamoeba species in human and animal hosts.

Whole genome sequencing of Entamoeba nuttalli reveals mammalian host-related molecular signatures and a novel octapeptide-repeat surface protein.

The enteric protozoa Entamoeba histolytica is the causative agent of amebiasis, which is one of the most common parasitic diseases in developed and developing countries. Entamoeba nuttalli is the genetically closest species to E. histolytica in current phylogenetic analyses of Entamoeba species, and is prevalent in wild macaques. Therefore, E. nuttalli may be a key organism in which to investigate molecules required for infection of human or non-human primates. To explore the molecular signatures of host-parasite interactions, we conducted de novo assembly of the E. nuttalli genome, utilizing self-correction of PacBio long reads and polishing corrected reads using Illumina short reads, followed by comparative genomic analysis with two other mammalian and a reptilian Entamoeba species. The final draft assembly of E. nuttalli included 395 contigs with a total length of approximately 23 Mb, and 9,647 predicted genes, of which 6,940 were conserved with E. histolytica. In addition, we found an E. histolytica-specific repeat known as ERE2 in the E. nuttalli genome. GO-term enrichment analysis of mammalian host-related molecules indicated diversification of transmembrane proteins, including AIG1 family and BspA-like proteins that may be involved in the host-parasite interaction. Furthermore, we identified an E. nuttalli-specific protein that contained 42 repeats of an octapeptide ([G,E]KPTDTPS). This protein was shown to be localized on the cell surface using immunofluorescence. Since many repeat-containing proteins in parasites play important roles in interactions with host cells, this unique octapeptide repeat-containing protein may be involved in colonization of E. nuttalli in the intestine of macaques. Overall, our draft assembly provides a valuable resource for studying Entamoeba evolution and host-parasite selection.

Endoparasitism of Greek ostriches: First report of Entamoeba struthionis and Balantioides coli.

Ostrich farming is a worldwide practice and an internationally developing industry. Among challenges in livestock production are the parasitic infections. The present study aimed to the investigation of the gastrointestinal parasites biota and prevalence in ostriches raised in different areas of Greece. A total of 141 clinically healthy ostriches originating from four different localities of Greece were coprologically examined for parasites of the gastrointestinal tract. Coprological examination revealed a considerably high rate of infection (65.9%) with protozoa; however, no helminths (trematodes, cestodes and nematodes) were detected. In detail, cysts of Entamoeba struthionis have been found in 57.4% of the examined birds. Moreover, 39.0% of sampled ostriches harboured cysts of Balantioides coli (syn. Balantidium coli), while oocysts of Cryptosporidium sp. were detected at a low percentage (2.1%). Partial sequences of the small subunit rRNA (16S rRNA) gene and the ITS region were amplified from pooled Entamoeba and Balantioides positive samples, respectively, confirming for the first time the presence of Entamoeba struthionis and Balantioides coli in ostriches in Greece. Some of these parasitoses require attention as they may affect productivity performance of the animals in commercial ostrich farming and possibly pose disease risk for livestock and humans.

Gastric and Caecal Amoebiasis in a Red Kangaroo (Macropus rufus) with Disseminated Toxoplasmosis.

A 1-year-old male red kangaroo (Macropus rufus) with an acute clinical history of lethargy, depression and increased respiratory rate was presented for necropsy examination. Gross lesions in the digestive tract were a distended stomach with watery content and multifocal raised ulcers covered by fibrinous pseudomembranes in the forestomach and caecum. On histopathology, there was necrotizing and ulcerative gastritis and typhlitis with intralesional amoebic trophozoites and Toxoplasma gondii zoites. Lesions due to T. gondii infection were observed in multiple organs and diagnosis was confirmed by fluorescent antibody test and immunohistochemistry. Both toxoplasmosis and gastric amoebiasis are diseases described in macropods. In this case report, we describe concurrent disease caused by both protozoa in a red kangaroo. Lesions of amoebiasis were also observed in the caecum. Both toxoplasmosis and amoebiasis should be considered as differential diagnoses of ulcerative lesions in the gastrointestinal tract in macropods during necropsy examination. Amoebiasis should be suspected especially when ulcerative lesions are observed in the forestomach.

Pathology Associated With an Outbreak of Entamoebiasis in Wild Cane Toads (Rhinella marina) in Tropical Australia.

Infection due to Entamoeba spp. is known to cause serious disease in primates (Entamoeba histolytica) and snakes (Entamoeba invadens), but there are no detailed descriptions of the pathology associated with Entamoeba spp. infection in amphibians. In 2014, an outbreak of entamoebiasis associated with a novel species of Entamoeba induced clinical illness and poor body condition in free-ranging cane toads in Australia's Northern Territory. Here, we describe the gross pathology, histology, and clinical pathology linked to the outbreak. The study compared 25 toads with invasive entamoebiasis, defined as histologically visible amoebas within tissue, and 12 toads without invasive entamoebiasis. Grossly, affected toads had mild to marked congestion of colonic serosal vasculature, with variable thickening of the intestinal wall and serosanguineous to hemorrhagic colonic content. Histologically, invasive entamoebiasis manifested primarily as moderate to severe, variably hyperplastic to ulcerative colitis. The small intestine was affected in 10 of 25 toads, and 5 of 25 toads also had gastric lesions. Amoebas consistent in morphology with Entamoeba sp. were commonly intermingled with mucosal epithelium, frequently along the basement membrane, with deeper invasion into the superficial lamina propria in only 5 toads. Toads with invasive entamoebiasis had neutrophilia, monocytosis, and lymphopenia, and thus elevated neutrophil to lymphocyte ratios, suggestive of an inflammatory and/or stress leukogram.

Molecular and descriptive epidemiology of intestinal protozoan parasites of children and their pets in Cauca, Colombia: a cross-sectional study.

BACKGROUND: Parasitic infections, particularly those caused by protozoa, represent a considerable public health problem in developing countries. Blastocystis, Giardia duodenalis, Cryptosporidium spp. and the Entamoeba complex (Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii) are the most common etiological causes of intestinal parasitic infections. METHODS: We carried out a descriptive cross-sectional study in school-age children attending a daycare institution in commune eight of Popayán, Cauca (Southwest Colombia). A total of 266 fecal samples were collected (258 from children and eight from pets). Blastocystis, G. duodenalis, Cryptosporidium spp. and the Entamoeba complex were identified by microscopy, quantitative real-time PCR (qPCR) and conventional PCR. The concordance of qPCR and microscopy was assessed using the Kappa index. Molecular characterization was conducted to identify Blastocystis subtypes (18S), G. duodenalis assemblages (tpi and gdh) and Cryptosporidium species/subtypes (18S and GP60). Potential associations between intestinal parasitism and sociodemographic factors were examined using bivariate analyses. RESULTS: A total of 258 fecal samples from children were analyzed by microscopy and 255 samples were analyzed by qPCR. The prevalence of Blastocystis was between 25.19% (microscopy) and 39.22% (qPCR), that of G. duodenalis was between 8.14% (microscopy) and 10.59% (qPCR), that of Cryptosporidium spp. was estimated at 9.8% (qPCR), and that of the Entamoeba complex was between 0.39% (conventional PCR) and 0.78% (microscopy). The concordance between microscopy and qPCR was very low. Blastocystis ST1 (alleles 4, 8, and 80), ST2 (alleles 11, 12, and 15), ST3 (alleles 31, 34, 36, 38,57, and 151), and ST4 (alleles 42 and 91), G. duodenalis assemblages AII, BIII, BIV and D, C. parvum subtype IIa and C. hominis subtype IbA9G3R2 were identified. The only identified member of the Entamoeba complex corresponded to E. histolytica. No statistically significant association was identified between parasitic infection and any sociodemographic variable. CONCLUSION: This study revealed the usefulness of molecular methods to depict the transmission dynamics of parasitic protozoa in southwest Colombia. The presence of some of these protozoa in domestic animals may be involved in their transmission.

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.