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Current status of intestinal parasite infections in fecal samples of dogs in Korea

You-Jeong LEE ; Beoul KIM ; Dongmi KWAK ; Min-Goo SEO

Parasites, Hosts and Diseases.2024;62(4):438-449. doi:10.3347/PHD.24024

This study surveyed the current status of intestinal parasite infections in Korean dog fecal samples. A total of 367 fecal samples were collected from the northern (Seoul and Gyeonggi-do), central (Chungcheong-do), and southern (Gyeongsang-do) regions and analyzed using the saturated sodium nitrate flotation technique and the nucleotide sequences of 18S rRNA. Six species of intestinal parasites were detected using the flotation technique. Among them, helminth eggs detected included Toxocara canis (6.0%), Toxascaris leonina (1.1%), Trichuris vulpis (6.8%), Ancylostoma caninum (2.7%), and Spirometra sp. (1.1%). Additionally, Cystoisospora sp. (7.6%) oocysts were also detected. The prevalence of intestinal parasite infections was higher in shelter dogs than in pet dogs. Molecular genetic assays revealed the gdh and 18S rRNA genes of Giardia duodenalis (type D) in 4.9% of fecal samples. To the best of our knowledge, 18S rRNA genes of Cryptosporidium canis were identified in 1.9% of fecal samples for the first time in Korea. These findings provide an overview of the current status of intestinal parasite infections in fecal samples of dogs from Korea and can be helpful in the surveillance of zoonotic parasite infections related to dogs.

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Probiotic-induced changes in intestinal microbiome inhibits Toxoplasma gondii infection

Hak-Jae LEE ; Do-Won HAM ; Seung-Hwan SEO ; Guang-Ho CHA ; Eun-Hee SHIN

Parasites, Hosts and Diseases.2024;62(4):408-423. doi:10.3347/PHD.24068

Toxoplasma gondii primarily invades the central nervous system, causing latent infections. Cysts persist in the host for life and there is currently no effective treatment. T. gondii infects human hosts through contaminated meat, invading the intestinal tissue and leading to changes in the number and composition of the gut microbiota. Since probiotic ingestion modulates intestinal microbiota changes, we hypothesized that intestinal microbiota dysbiosis caused by T. gondii infection would be restored following probiotic supplementation. To this end, we orally infected C57BL/6 mice with 10 T. gondii cysts and administered supplemental probiotics daily. We analyzed the levels of T. gondii B1 gene DNA, indicative of T. gondii infection, in brain tissue. We investigated alterations in the gut microbiota composition and functional pathways between the probiotic and non-probiotic treatment groups via next-generation sequencing analysis of each fecal sample. The infection level in the probiotic-treated group was significantly reduced after 4 weeks (p<0.05). Probiotic supplementation notably changed the gut microbiota after 2 weeks of infection, increasing the relative abundance of Intestinimonas massiliensis and Lawsonibacter asaccharolyticus. Probiotic supplements appear to modulate the gut microbiota, activating functional pathways involved in intestinal short-chain fatty acid production and strengthening the intestinal barrier, thereby impeding T. gondii infection and subsequent proliferation. Our findings provide valuable insights into T. gondii infection control and future study directions.

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Genotypic analysis of zoonotic Enterocytozoon bieneusi in wild deer in Korea

Gyeonguk NOH ; Haeseung LEE ; Seung-Hun LEE ; Min-Goo SEO ; Kyoo-Tae KIM ; Junho LEE ; Kaifa NAZIM ; Sang Joon PARK ; Man Hee RHEE ; Dongmi KWAK

Parasites, Hosts and Diseases.2024;62(4):484-489. doi:10.3347/PHD.24072

Enterocytozoon bieneusi is an important microsporidian protozoa that causes intestinal disorders in humans. We collected 191 fecal samples from roadkill deer carcasses, among which 13 (6.8%) showed positive reaction for E. bieneusi by polymerase chain reaction assay. Phylogenetic analysis revealed 6 distinct genotypes, 1 of which was novel. All genotypes belonged to Group 1, which has low host specificity, indicating possible transmission through sylvatic cycle. E. bieneusi infection was predominant in female deer (p<0.05).

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Microscopic findings of host–parasite interaction between the Korean rockfish, Sebastes schlegelii, and the monogenean ectoparasite, Microcotyle sebastis

Sang Phil SHIN ; Seokryel KIM

Parasites, Hosts and Diseases.2024;62(4):490-496. doi:10.3347/PHD.24043

This study was conducted to examine the pathogenesis of gill degeneration in the Korean rockfish Sebastes schlegelii, infested with the monogenean ectoparasite, Microcotyle sebastis. We collected 30 Korean rockfish from a fish farm in Tongyeong-si, Gyeongsangnam-do, and examined them with light microscopy, scanning electron microscopy (SEM), and histopathology, in March 2018. The monogenean trematode, M. sebastis, was detected in 27 Korean rockfish (90%), with the intensity of infection being 31.7 per fish. The characteristic surface ultrastructures such as tegument with transverse striations, genital atrium, genital pore, and opisthaptor with numerous clamps were observed. The worms were firmly attached to the gill lamellae using clamps from the opisthaptor, causing gill damage and degeneration. The distal part of the lamellae was ruptured by the sclerites of the clamps. The histopathological examination revealed epithelial hypertrophy, hyperplasia, and occasionally fusion of the lamellae. These ultrastructural and histopathological findings provide some understanding of the pathogenesis of gill degeneration in the Korean rockfish infested with M. sebastis.

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Clinical characteristics of toxoplasmosis patients in Korea: A retrospective study using health insurance review and assessment service data and electronic medical records

Do-Won HAM ; Bong-Kwang JUNG ; Ji-Hun SHIN ; Yong Joon KIM ; Kyoung Yul SEO ; Seung Mi LEE ; Jae Hyoung IM ; Jeong-Ran KWON ; Ho-Sung LEE ; Kyung-Won HWANG ; Eun-Hee SHIN

Parasites, Hosts and Diseases.2024;62(4):424-437. doi:10.3347/PHD.24001

This study aimed to elucidate the clinical characteristics of patients diagnosed with toxoplasmosis in Korea. We collected and analyzed the specific research data of 5,917 patients from the Health Insurance Review and Assessment (HIRA; 2007–2020) and 533 electronic medical records (EMRs; 2003–2021) of Korean patients. The HIRA data showed that toxoplasmosis is an endemic disease that occurs constantly in Korea, with a large proportion of patients complaining of ocular symptoms. Of the 533 patients for whom EMR data were available, 54.6% were diagnosed with toxoplasmosis; ocular toxoplasmosis (35.7%), congenital toxoplasmosis (4.7%), cerebral toxoplasmosis (4.1%), pulmonary toxoplasmosis (0.4%), and toxoplasma hepatitis (0.6%), in order of frequency. In ocular cases, 54.4% of the patients had diverse ocular pathologies. Toxoplasmosis in Korea is characterized by a high frequency of ocular symptoms, most patients are adults, and 51.8% of patients with seropositivity were positive for IgG, suggesting prior infection. This study highlights that patients with ocular symptoms are included in the major diagnosis group for acquired toxoplasmosis in Korea.

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Functional characterization of glucose transporter 4 involved in glucose uptake in Clonorchis sinensis

Hojong JUN ; Ernest MAZIGO ; Wang-Jong LEE ; Yun-Kyu PARK ; Jin-Hee HAN ; Seok Ho CHA

Parasites, Hosts and Diseases.2024;62(4):450-460. doi:10.3347/PHD.24051

Clonorchis sinensis, which causes clonorchiasis, is prevalent in East Asian countries and poses notable health risks, including bile duct complications. Although praziquantel is the primary treatment for the disease, the emerging resistance among trematodes highlights the need for alternative strategies. Understanding the nutrient uptake mechanisms in trematodes, including C. sinensis, is crucial for developing future effective treatments. This study aimed to characterize the function of C. sinensis glucose transporter 4 (CsGTP4) and determine its role in nutrient uptake employing synthesized cDNA of adult C. sinensis worms. The functional characterization of CsGTP4 involved injecting its cRNA into Xenopus laevis oocytes and analyzing the deoxy-D-glucose uptake levels. The results demonstrated that deoxy-D-glucose uptake depended on the deoxy-D-glucose incubation and CsGTP4 expression time, but not sodium-dependent. The concentration-dependent uptake followed the Michaelis–Menten equation, with a Km value of 2.7 mM and a Vmax value of 476 pmol/oocyte/h based on the Lineweaver–Burk analysis. No uptake of radiolabeled α-ketoglutarate, p-aminohippurate, taurocholate, arginine, or carnitine was observed. The uptake of deoxy-D-glucose by CsGTP4 was significantly inhibited by unlabeled glucose and galactose in a concentration-dependent manner. It was significantly inhibited under strongly acidic and basic conditions. These insights into the glucose uptake kinetics and pH dependency of CsGTP4 provide a deeper understanding of nutrient acquisition in trematodes. This study contributes to the development of novel antiparasitic agents, addressing a considerable socioeconomic challenge in affected regions.

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Allyl isothiocyanate exacerbates acute toxoplasmosis through inhibition of inflammatory cytokines

Qiu-Mei LIN ; Hong-Bin LONG ; Jun-Ting HE ; Zhi-hao ZHANG ; Ho-Woo NAM ; Fu-Shi QUAN ; Qi ZHONG ; Xu-Qing LIU ; Zhao-Shou YANG

Parasites, Hosts and Diseases.2024;62(4):476-483. doi:10.3347/PHD.24054

Allyl isothiocyanate (AITC) is a natural product commonly used in food preservation and pharmaceutical applications. Toxoplasmosis, caused by the protozoan pathogen Toxoplasma gondii, is prevalent globally while the impact of AITC on toxoplasmosis is unclear. We explored the effect of AITC on acute toxoplasmosis. We infected C57BL/6 mice with T. gondii type I RH strain following AITC administration. On the 4th day after infection, which corresponds to the initial stage of infection, we collected serum for the determination of inflammatory cytokine levels. The mice serum of the AITC-administered group contained significantly lower levels of granulocyte colony-stimulating factor, interferon-gamma, interleukin (IL)-23 subunit p19, IL-4, IL-6, and monocyte chemoattractant protein-1. The lifespan of the mice in the AITC-administered group was significantly reduced. In vitro experiments showed that AITC promoted the proliferation of intracellular T. gondii accompanied by the inhibition of IL-4, IL-1β, and IL-6 production in RAW264.7 macrophages. Our results showed that AITC facilitated T. gondii infection in the early stage by inhibiting the production of several inflammatory cytokines.

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Identification of essential genes for Acanthamoeba castellanii excystation during encystation and excystation

Min-Jeong KIM ; Hye-Jeong JO ; Fu-Shi QUAN ; Ki Back CHU ; Hyun-Hee KONG ; Eun-Kyung MOON

Parasites, Hosts and Diseases.2024;62(4):399-407. doi:10.3347/PHD.24062

Acanthamoeba is an opportunistic pathogen that causes Acanthamoeba keratitis, granulomatous amoebic encephalitis, and other cutaneous diseases. The life cycle of Acanthamoeba consists of 2 stages of trophozoites and cysts. Under adverse environmental conditions, Acanthamoeba encysts, while the conditions become favorable for growth, it reverts to the trophozoite form. Acanthamoeba excystation is crucial for its proliferation and can lead to recurrent infections after incomplete treatment. To identify the factors involved in excystation, A. castellanii was subjected to either encystation- or excystation-inducing conditions, and gene expression profiles were compared using mRNA sequencing. A. castellanii samples were collected at 8 h intervals for analysis under both conditions. Differentially expressed gene analysis revealed that 1,214 and 1,163 genes were upregulated and downregulated, respectively, by more than 2-fold during early excystation. Five genes markedly upregulated in early excystation (ACA1_031140, ACA1_032330, ACA1_374400, ACA1_275740, and ACA1_112650) were selected, and their expression levels were confirmed via real-time PCR. Small interfering RNA (siRNA) targeting these 5 genes was transfected into Acanthamoeba and gene knockdown was validated through real-time PCR. The silencing of ACA1_031140, ACA1_032330, ACA1_374400, and ACA1_112650 inhibited excystation and suggested that these genes might be essential for excystation. Our findings provide valuable insights for suppressing Acanthamoeba proliferation and recurrence.

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Understanding the pathogenicity of Naegleria fowleri in association with N. fowleri antigen-1 (Nfa1)

Jong-Hyun KIM ; Hae-Jin SOHN ; Ho-Joon SHIN ; Stacy E. WALZ ; Suk-Yul JUNG

Parasites, Hosts and Diseases.2024;62(4):385-398. doi:10.3347/PHD.24025

Naegleria fowleri, a brain-eating amoeba, thrives in lakes and rivers with aquatic vegetation and causes primary amoebic meningoencephalitis (PAM) in humans. Most recently, it has become such a serious problem that N. fowleri was detected in tap water in Houston, USA. Several pathogenic factors are considered very important to destroy target cells in the brain. In particular, the food-cup where N. fowleri antigen-1 (Nfa1) is located, is strongly expressed in pseudopodia involved in the movement of N. fowleri, and is involved in phagocytosis by attaching to target cells. In this article, we reviewed the role of the Nfa1 protein and its associated pathogenicity. The nfa1 gene was cloned by cDNA library immunoscreening using infection serum and immune serum. Nfa1 protein is mainly distributed in pseudopodia important to movement and vacuoles. Moreover, heat shock protein 70, cathepsin-like proteare and Nf-actin are also associated with pseudopodia in which Nfa1 is localized. Interestingly, the amount of the nfa1 gene changed as N. fowleri trophozoites transformed into cysts. Polyclonal antiserum against Nfa1 showed a protective effect against cytotoxicity of approximately 19.7%. Nfa1-specific IgA antibodies prevent N. fowleri trophozoites from adhering to the nasal mucosa, delaying invasion. The nfa1-vaccinated mice showed significantly higher levels of Nfa1-specific antibody. The duration of anti-Nfa1 IgG in the vaccinated mice lasted 12 weeks, strongly suggesting that nfa1 is a significant pathogenic gene and that Nfa1 is a pathogenic protein. Several factors related to pseudopodia and locomotion have been linked to Nfa1. A clearer function of N. fowleri targeting nfa1 with other genes might enable target-based inhibition of N. fowleri pathogenicity.

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Infection status and species diversity of trematode cercariae in freshwater snails from canal networks in the Bangkok Metropolitan Region, Thailand

Phuphitchan RACHPRAKHON ; Watchariya PURIVIROJKUL

Parasites, Hosts and Diseases.2024;62(4):461-475. doi:10.3347/PHD.24033

This study aimed to investigate the infection status and species diversity of trematode cercariae in freshwater snails from canal networks in the Bangkok Metropolitan Region (BMR), Thailand. The snails were collected from 35 sites during 2 cycles of the wet (July–October) and dry (November–June) seasons in 2018–2019. A total of 29,420 snails representing 24 species/subspecies were examined for cercarial infection using shedding and crushing techniques. We found that 1,275 snails from 12 species/subspecies were infected, resulting in an overall prevalence of 4.3%. Infections were significantly higher (p<0.001) during the wet season (5.9%; 970/16,473) than during the dry season (2.4%; 305/12,947). Morphological identification revealed 37 distinct types within 15 morphotypes, with the armatae morphotype showing the highest infection rate (1.8%) and the greatest cercarial diversity (8 distinct types). This study highlights the health risk posed by the Bithynia siamensis siamensis, which was the second most prevalent (8.5%) and hosted the greatest cercarial diversity (11 morphotypes, 15 distinct types). This subspecies also displayed a wide distribution range (31 localities) with a generally high occurrence frequency in the BMR. This study firstly documents a gymnophallid digenean as a freshwater digenean, presenting evidence of a dichotoma cercarial morphotype from 4 species/subspecies (Filopaludina martensi martensi, F. sumatrensis polygramma, B. siamensis siamensis, and Wattebledia siamensis) with a low infection range (0.1–0.4%). A staggering diversity of cercariae was observed in the BMR canal networks with seasonal fluctuations. The B. siamensis siamensis displayed notable epidemiological importance in the BMR flowing-water networks. This study provides quantitative and qualitative morphological descriptions and measurement guidelines for the dichotoma cercaria in Thailand.

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