Immunohistochemical detection and pathology of toxoplasmosis in Nigerian wild rats.

Toxoplasmosis is a zoonotic disease of economic importance found worldwide, and it is caused by Toxoplasma gondii, which affects a wide range of hosts. High prevalence of toxoplasmosis has been reported in rodents, and they are considered very important in the circulation and maintenance of the disease. However, epidemiologic studies of the disease in rodents are generally scarce in the Tropics. This study utilized the immunohistochemical (IHC) technique to detect Toxoplasma gondii in wild rats sampled from across the North Central Nigeria. The brain, intestine, diaphragm, lungs and kidney tissue samples from 227 wild rats (Zyzomys pedunculatus) were routinely processed for histopathology, out of which 86 were further selected for IHC detection of T. gondii antigens using the streptavidin-peroxidase method. The histologic lesions observed were mild to moderate in severity, including meningitis, focal gliosis, neuronal degeneration and necrosis, villous atrophy and denudation, enteritis, diaphragmatic myositis, broncho-interstitial pneumonia and interstitial nephritis. Toxoplasma gondii was detected in 82.6% of the selected samples showing various degrees of immunoreaction intensity. We conclude that IHC is a useful tool in the detection of T. gondii in wild rats, and lungs and kidney may be the organ of choice for the detection of T. gondii.

Novel genotypes and multilocus genotypes of Enterocytozoon bieneusi in two wild rat species in China: potential for zoonotic transmission.

Enterocytozoon bieneusi is an opportunistic pathogen in immunodeficient patients. Although this pathogen has been reported in many domestic animals, few data are available about the occurrence of E. bieneusi in wild rats. In the current study, a total of 228 fecal samples from two wild rat species (Leopoldamys edwardsi and Berylmys bowersi) in China were examined by a nested PCR-based sequencing approach employing the internal transcribed spacer (ITS) region of nuclear ribosomal DNA. The overall prevalence of E. bieneusi in wild rats was 33.3% (76/228), with 35.1% (39/111) in L. edwardsi and 31.6% (37/117) in B. bowersi. Ten E. bieneusi genotypes (including four known and six novel genotypes) were identified, with the novel CQR-2 (n = 15) as the predominant genotype. Phylogenetic analysis indicated that ten genotypes in the present study belong to zoonotic group 1, which contains many genotypes in humans. Furthermore, multilocus sequence typing (MLST) analysis showed that 19 ITS-positive samples were successfully amplified at three microsatellites and one minisatellite, forming 18 multilocus genotypes (MLGs). This is the first report of E. bieneusi infection in the wild rats L. edwardsi and B. bowersi. Our findings suggest that wild rats could be a significant source of human infection, including contaminated food and water.

Molecular Detection and Genotyping of Toxoplasma gondii in Edward's Long-Tailed Rats (Leopoldamys edwardsi).

Toxoplasma gondii is an important zoonotic parasite infecting humans and various animals with a worldwide distribution. However, limited information is available on T. gondii infection in wild rats. The present study aimed to examine the prevalence and characterize the genotypes of T. gondii in wild rats in two regions of China. Brain tissues were collected from 111 Edward's long-tailed rats (Leopoldamys edwardsi) and 117 Bower's white-toothed rats (Berylmys bowersi) between November 2017 and January 2018. Genomic DNA was extracted and amplified by PCR targeting the T. gondii B1 gene. B1 gene-positive samples were genotyped at 10 genetic markers (SAG1, SAG2 [5', 3'] and [alternative], SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico) using multilocus nested polymerase chain reaction/restriction fragment length polymorphism. Six (5.41%, 6/111) Edward's long-tailed rats from Chongqing Municipality were positive for T. gondii B1 gene, whereas no T. gondii infection was detected in Bower's white-toothed rats (n = 117) from Guangdong province. T. gondii prevalence in female and male rats was 1.77% (2/113) and 3.48 (4/115), respectively. Four of the six positive DNA samples were completely genotyped at 10 genetic loci and were identified as ToxoDB#20. The present study revealed the occurrence of T. gondii infection in Edward's long-tailed rats. These findings raised public health concerning about T. gondii infection in wild rats. These results provide reference data for understanding the distribution of T. gondii genotypes in wild rats in China.

Bufavirus Protoparvovirus in feces of wild rats in China.

Bufavirus (BuV) was first discovered from feces of children with acute diarrhea. It was subsequently detected from several animal species including shrews, bats, and nonhuman primates. In this study, we identified a novel Protoparvovirus, designated RatBuV, from the intestinal contents of wild rats using viral metagenomics. The near complete genome was 4643 nt encoding NS1, VP1, and VP2 proteins. Phylogenetic analysis over the complete genome showed that RatBuV clustered with Mpulungu BuV from shrews. Sequence analysis indicated that the putative protein sequences of NS1, VP1, and VP2 of RatBuV shared identities of 50.6-77.2, 48.3-77.3, and 47.1-78.3 %, respectively, with those of human BuVs, MpBuV, and WUHARV parvovirus, suggesting RatBuV belongs to a new species of Protoparvovirus. Our epidemiologic study indicated that the prevalence rate of RatBuV in the cohort of 40 wild rats is 12.5 % (5/40), which is higher than that of BuV in humans in a previous study.

Identification of rickettsiae from wild rats and cat fleas in Malaysia.

Rickettsioses are emerging zoonotic diseases reported worldwide. In spite of the serological evidence of spotted fever group rickettsioses in febrile patients in Malaysia, limited studies have been conducted to identify the animal reservoirs and vectors of rickettsioses. This study investigated the presence of rickettsiae in the tissue homogenates of 95 wild rats and 589 animal ectoparasites. Using PCR assays targeting the citrate synthase gene (gltA), rickettsial DNA was detected in the tissue homogenates of 13 (13.7%) wild rats. Sequence analysis of the gltA amplicons showed 98.6-100% similarity with those of Rickettsia honei/R. conorii/R. raoultii (Rickettsiales: Rickettsiaceae). Sequence analysis of outer membrane protein A gene (ompA) identified Rickettsia sp. TCM1 strain from two rats. No rickettsia was detected from Laelaps mites, Rhipicephalus sanguineus and Haemaphysalis bispinosa ticks, and Felicola subrostratus lice in this study. R. felis was identified from 32.2% of 177 Ctenocephalides felis fleas. Sequence analysis of the gltA amplicons revealed two genotypes of R. felis (Rf31 and RF2125) in the fleas. As wild rats and cat fleas play an important role in the enzoonotic maintenance of rickettsiae, control of rodent and flea populations may be able to reduce transmission of rickettsioses in the local setting.

Molecular detection of toxoplasmosis in wild rats using loop-mediated isothermal amplification assay.

Background and Aim: Toxoplasmosis is caused by the parasite Toxoplasma gondii. Cats are the only known hosts that excrete resistant oocysts. Wild rats serve as crucial reservoirs and intermediate hosts for T. gondii's survival and dissemination. Consuming soil and water containing oocysts can lead to illness. This study aimed to estimate the prevalence of toxoplasmosis in wild rats through molecular detection as an indicator of environmental contamination in Surabaya. Materials and Methods: One hundred rats were collected from the three areas (housing, dense settlements, and traditional markets) and distributed into the five zones: West, East, Central, North, and South of Surabaya. Brain tissue samples were extracted using a Geneaid™ (New Taipei City, Taiwan) DNA isolation kit and analyzed through the loop-mediated isothermal amplification (LAMP) method. Results: The study analyzed brain tissue from 100 wild rats, consisting of 77 Rattus tanezumi and 33 Rattus norvegicus, displaying 30% LAMP positivity. The study revealed that 30% (30/100) of wild rats tested were infected with T. gondii. The molecular prevalence rate in male rats was 32.35% (22/68), compared to females with 25% (8/32). 41.9% of the housing population, 33.3% of traditional markets, and 22.6% of dense settlements had the highest molecular prevalence. The high positive molecular rate at the trapping site can be attributed to cats and dense populations. Conclusion: Thirty percentage wild rats were tested positive for toxoplasmosis in Surabaya, East Java, Indonesia using LAMP method. Implementing strict control and monitoring is crucial in preventing the transmission of diseases from wild rats to humans. It is necessary to carry out further research related to genetic analysis of T. gondii to determine the type of T. gondii that infects animals and humans in Surabaya through bioassay and molecular test.

Tissue cysts and serological detection toxoplasmosis among wild rats from Surabaya, East Java, Indonesia.

Background: The protozoan Toxoplasma gondii is the source of zoonosis toxoplasmosis and causes public health problems throughout the world. Environmental contamination by oocysts excreted by cats as definitive hosts affects the spread of this disease. Wild rats as rodents can be used as an indicator of environmental contamination by oocysts, considering that rats have a habit of living in dirty environments and can be infected by oocysts from the environment. Aim: This study aims to detect toxoplasmosis from tissue cysts and serological tests in wild rats as an indicator of environmental contamination in Surabaya. Methods: A total of 100 wild rats collected from Surabaya were collected in five areas (West, East, Central, North, and South of Surabaya) obtained from three trapping locations: housing, dense settlements, and markets. All samples were examined microscopically for parasitological tests through the brain tissue samples, and the serum was examined using the toxoplasma modified agglutination test to detect the presence of IgG and Immunoglobulin M (IgM). Results: This research used 100 wild rat samples, 77 Rattus tanezumi and 33 Rattus norvegicus, with evidence of 31% in serology and active infection with 19% tissue cyst. The results showed that the seroprevalence of T. gondii in wild rats was 31% (30% for IgG and 1% for IgM). Tissue cysts in the rat brain samples tested were 19% (19/100). The IgG prevalence rate in female rats was 25% (8/32), while for males, it was 32.3% (22/68). The highest seropositive IgG from densely populated settlements was 50%, markets were 25.8%, and housing was 12.1%. The highest seropositive IgM from densely populated settlements was 2.8%. Population density and the presence of cats are factors supporting the high seropositive rate at the trapping location. Conclusion: This study revealed that there has been toxoplasmosis contamination in Surabaya with evidence of 31% in serology and active infection with 19% tissue cyst. It is necessary for controlling with surveillance in cats to prevent transmission in humans.

High prevalence of Toxoplasma gondii in Nigerian wild rats by molecular detection.

Toxoplasmosis has been reported in Nigeria using several diagnostic tools with high prevalence in humans and some food animals. Rodents have been recognised as vital intermediate hosts of Toxoplasma gondii. However, there is paucity of information on the occurrence of T. gondii in wild rats found in Nigeria. This study aimed at molecular detection of T. gondii in Zyzomys pedunculatus and to evaluate its involvement in the epidemiology of toxoplasmosis in Nigeria. A total of 84 rats were sampled across three states of the North Central Nigeria, and DNA was extracted from the brain, lungs, kidney and intestine of the rats for the detection of T. gondii DNA by nested PCR to amplify the multicopy B1 gene. Sixty-four of the 84 samples (76.2%) were positive for T. gondii out of which 5 samples were sequenced and had an identity score of between 97.73% and 99.35% with the reference B1 gene of T. gondii in GenBank. This study suggests Nigerian wild rats may be an important intermediate hosts of T. gondii and may play a role in the epidemiology and maintenance of T. gondii circulation in Nigeria.

A global overview of the most important zoonotic bacteria pathogens transmitted from Rattus norvegicus to humans in urban environments.

Zoonotic pathogens, comprising over 61% of all pathogenic microorganisms, can be transmitted from different animals to individuals in numerous ways either in the presence or the absence of a vector. Causing new emerging human infectious diseases, these pathogens could be categorized into 4 groups, bacteria, viruses, parasites, and fungi. Among the wide range of reservoirs for zoonotic pathogens, tremendous attention has been attracted to wild rats, due to their global distribution not only in urban environments but also in the sylvatic and agricultural surroundings. For the nonce, zoonotic bacteria transmitted via wild rats have turned into a global public health problem probably due to their ability to induce re-emerging diseases even after eradication and controlling management. Despite the importance of wild rats in spreading pathogens, little data are available about the bacterial diversity present in urban wild rat populations. In this review, we present a complete list of zoonotic bacterial pathogens isolated from wild rats in urban environments.

The Detection of Toxoplasma gondii in Wild Rats (Rattus norvegicus) on Mink Farms in Shandong Province, Eastern China.

Toxoplasma gondii is a worldwide distributed zoonotic pathogen that threatens public health. However, there have been limited data for T. gondii infection in wild rats (Rattus norvegicus) in China. In the present study, a total of 227 wild rats were captured from three mink farms to investigate T. gondii infection in Shandong Province, eastern China. The DNA was extracted from 25 mg rats' brain tissues and subjected to a PCR amplification by targeting to the T. gondii B1. In 227 wild rat samples, 18 samples (7.93%) were positive for T. gondii. Then, the positive samples were further genotyped based on eight genetic markers, including eight nuclear loci (SAG1, 5'-SAG2, and 3'-SAG2, alternative SAG2, SAG3, GRA6, c29-2, and L358) and an apicoplast locus (Apico) by using the multilocus PCR-restriction fragment length polymorphism technology. Of these samples, eight were genotyped at nine nuclear loci, and two were genotyped at eight nuclear loci, forming three known genotypes (ToxoDB no. 43, ToxoDB no. 91, and ToxoDB no. 189) and two new genotypes. The closest ToxoDB genotypes were observed in wild rats, suggesting the differences in the population structure of the T. gondii between breed farm animals and wild rats. These data revealed the genetic variability of T. gondii in wild rats on mink farms in Shandong Province, with possible implication for public health.

First detection and characterization of rat hepatitis E Virus (HEV-C1) in Japan.

Rat hepatitis E virus (HEV-C1) in the Orthohepevirus C species has been reported to cause zoonotic infection and hepatitis in humans. HEV-C1 strains have been detected from wild rats in many countries in Europe, Asia, and North America. However, in Japan, no HEV-C1 strains have been identified. In the present study, 5 (1.2%) of 428 wild rats (Rattus norvegicus or R. rattus) were positive for anti-HEV-C1 IgG. Although all 428 rat sera were negative for HEV-C1 RNA, it was detectable in 20 (19.8%) of 101 rat fecal samples collected on a swine farm, where HEV (genotype 3b, HEV-3b) was prevalent and wild rats were present. In addition, HEV-C1 RNA was detectable in the intestinal contents and liver tissues of 7 (18.9%) of 37 additional rats captured on the same farm. The HEV-C1 strain (ratEJM1703495L) obtained in this study shared only 75.8-84.7% identity with reported HEV-C1 strains over the entire genome but propagated efficiently in cultured cells. HEV-3b strains were detected in the rats' intestinal contents, with 97.3-99.5% identity to those in pigs on the same farm, but were undetectable in rat liver tissues, suggesting that wild rats do not support the replication of HEV-3b of swine origin.

Prevalence of Leptospira interrogans in wild rats (Rattus norvegicus and Cricetomys gambianus) in Zaria, Nigeria.

Leptospirosis is a neglected disease of zoonotic importance and rodents have a known role in epidemiology of Leptospira globally. Paucity of information on the prevalence of leptospirosis in wild rats used as games in Zaria, Nigeria informed the study. The study aimed to detect Leptospira interrogans in wild rats in Zaria, Nigeria. A total of 71 wild rats comprising 57 Rattus norvegicus and 14 Cricetomys gambianus were sampled over a period of 3 months (April-June 2019). Fisher exact test was used with confidence interval set at 0.05 to ascertain associations between positive cases and species. Blood was collected from 56 rats and harvested sera screened for Leptospira interrogans antibody using rat IgG competitive enzyme linked immunosorbent assay (c-ELISA). Following humane euthanasia of rats, 71 samples (62 kidney tissues and 9 urine samples) were collected in sterile labeled tubes and cultured using Ellinghausen Mc-cullough Johnson Harris (EMJH) enrichment and basal medium. Results indicated over all Leptospira spp antibody detection of 73.2 % (41/56) in Rattus norvegicus (60.7 %) and Cricetomys gambianus (12.5 %). No significant difference (P > 0.05) existed for the prevalence of Leptospira interrogans antibody in the species of wild rats. Over all occurrence of Leptospira interrogans were 74.2 % (46/62) in kidneys and 55.6 % (5/9) in urine samples. Based on species of rats, Rattus norvegicus recorded prevalence of 76.9 % (40/52) and 40.0 % (2/5) in kidney and urine samples respectively. Prevalence of 60.0 % (6/10) and 75.0 % (3/4) in kidney and urine samples respectively were recorded for Cricetomys gambianus. There was significant difference (P < 0.05) in the prevalence of Leptospira interrogans in kidney samples of both wild rats. These species of rats could be reservoirs of Leptospira interrogans. The result showed high prevalence of Leptospira spp in the wild rats and the possibility of domestic animals and humans contracting the disease. This study is the first documentation of evidence of pathogenic Leptospira species in wildlife used as games in Zaria, Nigeria.

Molecular Detection of Cryptosporidium spp. and Enterocytozoon bieneusi Infection in Wild Rodents From Six Provinces in China.

Enterocytozoon (E.) bieneusi and Cryptosporidium spp. are the most important zoonotic enteric pathogens associated with diarrheal diseases in animals and humans. However, it is still not known whether E. bieneusi and Cryptosporidium spp. are carried by wild rodents in Shanxi, Guangxi, Zhejiang, Shandong, and Inner Mongolia, China. In the present study, a total of 536 feces samples were collected from Rattus (R.) norvegicus, Mus musculus, Spermophilus (S.) dauricus, and Lasiopodomys brandti in six provinces of China, and were detected by PCR amplification of the SSU rRNA gene of Cryptosporidium spp. and ITS gene of E. bieneusi from June 2017 to November 2020. Among 536 wild rodents, 62 (11.6%) and 18 (3.4%) samples were detected as E. bieneusi- and Cryptosporidium spp.-positive, respectively. Differential prevalence rates of E. bieneusi and Cryptosporidium spp. were found in different regions. E. bieneusi was more prevalent in R. norvegicus, whereas Cryptosporidium spp. was more frequently identified in S. dauricus. Sequence analysis indicated that three known Cryptosporidium species/genotypes (Cryptosporidium viatorum, Cryptosporidium felis, and Cryptosporidium sp. rat genotype II/III) and two uncertain Cryptosporidium species (Cryptosporidium sp. novel1 and Cryptosporidium sp. novel2) were present in the investigated wild rodents. Meanwhile, 5 known E. bieneusi genotypes (XJP-II, EbpC, EbpA, D, and NCF7) and 11 novel E. bieneusi genotypes (ZJR1 to ZJR7, GXM1, HLJC1, HLJC2, and SDR1) were also observed. This is the first report for existence of E. bieneusi and Cryptosporidium spp. in wild rodents in Shanxi, Guangxi, Zhejiang, and Shandong, China. The present study also demonstrated the existence of E. bieneusi and Cryptosporidium spp. in S. dauricus worldwide for the first time. This study not only provided the basic data for the distribution of E. bieneusi and Cryptosporidium genotypes/species, but also expanded the host range of the two parasites. Moreover, the zoonotic E. bieneusi and Cryptosporidium species/genotypes were identified in the present study, suggesting wild rodents are a potential source of human infections.

Identification of Cryptosporidium viatorum XVa subtype family in two wild rat species in China.

BACKGROUND: Cryptosporidium viatorum is a minor Cryptosporidium pathogen in humans. Currently, there is limited information regarding the prevalence and genotypes of C. viatorum in animals in China. METHODS: In this study, 228 faecal samples were collected from two wild rat species (Leopoldamys edwardsi and Berylmys bowersi) in Chongqing Municipality and Guangdong Province, China. These specimens were analyzed for C. viatorum and then subtyped it using PCR and sequence analysis of the small subunit ribosomal RNA (SSU rRNA) and 60-kilodalton glycoprotein (gp60) genes, respectively. RESULTS: A total of 25 (11.0%) faecal samples were tested positive for C. viatorum by SSU rRNA assay. Of these samples, 4 (3.6%) came from L. edwardsi and 21 (18.0%) from B. bowersi. Of the 25 C. viatorum-positive samples, 17 were successfully amplified at the gp60 gene locus, which represented four subtypes belonging to two subtype families, including XVa (XVaA6, XVaA3g, XVaA3h) and XVc (XVcA2G1). Phylogenetic analysis based on the gp60 amino acid sequences indicated that all of the C. viatorum isolates grouped together, supporting the conclusion that C. viatorum from the wild rats represent two subtype families. CONCLUSIONS: These results indicate an occurrence of C. viatorum XVa subtype family from rats which is genetically identical to those found in humans. Our findings suggest that wild rats may be a potential source of human cryptosporidiosis.

Cysticercus fasciolaris infection in wild rats (Rattus norvegicus) in Korea and formation of cysts by remodeling of collagen fibers.

Cysticercus fasciolaris, the larval form of Taenia taeniaeformis, is commonly encountered in rodents. In our study, 287 wild rats (Rattus norvegicus) in South Korea were examined in 2010 and 2011. Of 287 rats, 97 (33.8%) were infected with C. fasciolaris A strong positive correlation was found between the host body weight and prevalence in both sexes, regardless of the year of collection. The liver was the most common habitat of the parasite, and the lung was the most frequent ectopic region, followed by mesentery, pleura, abdominal wall, and kidney. The lesions of the affected organs were generally characterized by well-developed cysts, each containing a larva. However, the cysts within kidney and abdominal wall were poorly organized, filled with abscess, and lacked larvae. Collagen types I and III, but not type IV, played significant roles in constructing the cysts at differential stages, addressed by immunohistochemistry. During cyst wall development, both collagen types contributed equally to cyst formation at the early stage, whereas collagen type I was the major component at the late stage (p < 0.05). In early-stage cysts, distribution of collagens was interestingly differential depending on the development stage, as collagen type I was localized in the outer layer and type III was located in the inner layer. Our results suggest that an appropriate remodeling process of collagen fibers is necessary for C. fasciolaris to build the well-conditioned cysts in the target organs for survival.

Nota sopre uma espécie de Hepatozoon (Protozoa, Haemogregarinidae), parasita de um rato silvestre do gênero Zygodontomys, de São Paulo, Brasil / Note on a species of Hepatozoon (Protozoa, Haemogregarinidae), parasite of a wild rat of the genus Zygodontomys, from São Paulo, Brazil

Os autores descreveram uma espécie de Hepatozoon em rato silvestre Zygodontomys sp., encontrado nas células da medula óssea. A espécie é diferente de outra anteriormente descrita em Oryzomys sp., porém os autores não lhe dão nome específico, esperando para isso o estudo de sua evolução (AU).