Report Of Natural Infection Research Articles

Pathological and molecular characteristics of a new infectious Streptococcus agalactiae from farmed Schizopygopsis pylzovi in southern China

During the months of November and December 2022, a severe disease outbreak led to substantial mortality among Schizopygopsis pylzovi (S. pylzovi) in the Manwan reservoir of Lincang, China. The prominent clinical symptoms observed were exophthalmos with cataract, iris congestion, splenomegaly, and a pale liver. To elucidate the etiology, bacterial isolation was performed, followed by identification using Diff-Quik staining, 16s rDNA sequencing, and specific PCR. The results revealed that all three isolates were a non-hemolytic streptococcus with 93% identity to the 16S rDNA sequence of Streptococcus agalactiae (S. agalactiae), and further specific PCR demonstrated that it belonged to S. agalactiae. In addition, to further elucidate the molecular characteristics of the pathogen, a comprehensive analysis was conducted, encompassing pathological diagnosis, serotyping, multilocus sequence typing (MLST), physiological and biochemical identification, as well as the detection of related virulence genes. The findings showed that the isolates were S. agalactiae serotype Ib ST-891, exhibiting significant damage to the eye, spleen, and brain. Notably, its virulence gene profile and physiological and biochemical characteristics exhibited a significant degree of similarity to those from S. agalactiae serotype Ib ST261. Finally, experimental infection revealed that mortality rates were 100% and 75%, respectively, after exposure to 1 × 108 CFU/mL and 1 × 107 CFU/mL of this isolate suspension. It is the first report of natural infection with S. agalactiae serotype Ib ST-891 in fish, thus providing evidence for the molecular genetic diversity of fish-derived S. agalactiae.

First report of alstroemeria mosaic virus infecting Alstroemeria in Korea.

Alstroemeria, a member of the Alstroemriaceae family, is a popular cut flower plant with a long-base life and a wide variety of flower colors. It is widely cultivated in many countries, especially in Central and South America. However, numerous viruses such as alstroemeria carlavirus (AlCV), alstroemeria mosaic virus (AlMV), cucumber mosaic virus (CMV), tomato spotted wilt virus (TSWV), alstroemeria streak virus (AlSV), and impatiens necrotic virus (INSV) can infect Alstroemeria and significantly decrease its yield (Kim, 2020). Among these viruses, AlMV is well known to cause an endemic viral disease in the Netherlands (Corine M. et al. 1992). AlMV is a member of the genus potyvirus in the family Potyviridae, one of the most widely distributed families of plant viruses. In 2021, symptomatic alstroemeria plants showing interveinal leaf streaking with elongated light green and chlorosis of leaves were identified from farms in a greenhouse in Gwangju, South Korea. Potyvirus-like particles (approximately 750-800 nm in length) were observed from sap of the symptomatic plants by electron microscope (Supplementary Fig. 1). To confirm virus infection, total RNA was extracted from an alstroemeria leaf using a Beniprep® Super Plant RNA extraction kit (IVT7005, Invirustech Co., Korea). A cDNA library was synthesized and analyzed by high throughput sequencing (HTS) using an Illumina NovaSeq6000 S4 sequencer. A total of 48,072,240 raw reads were obtained after quality filtering with FastQC. Remaining sequences were de novo assembled into contigs with a Trinity assembler. Nucleotide blast analysis of contigs against NCBI viral reference database revealed that 24 assembled contigs (> 1,000 bp) were sequences of AlMV. To confirm AlMV detection, raw reads were mapped to known AlMV complete genome (9,774 bp) using Bowtie2 program. Results showed that a total of 4,698,112 reads were mapped. A consensus sequence (9,778 bp, accession no. LC709275) was then obtained. To verify the presence of AlMV, RT-PCR assay was conducted with AlMV's CP gene-specific primers: AlMV-F (5'-CACGAGGCTGTGAAACAAGC -3') and AlMV-R (5'- CCAGGCGACACGGCTAAATA-3'). PCR products of the expected size (538 bp) were cloned, sequenced, and subjected to GenBank BLASTn search. A 538 bp partial CP sequence was used for BLAST analysis which revealed that it shared 100% identities with the consensus sequence (LC709275) and 96.99~98.76% nucleotide identities with four AlMV isolates (MK440140, NC043135, MT892648, DQ295032). Phylogenetic analysis based on partial CP sequences of representative members of potyviruses (family Potyviridae) using 1,000 bootstrap replicates based on either neighbor-joining or Kimura 2 parameter methods in MEGA-X revealed that AlMV isolate JNU-2 was grouped together with the four known AlMV isolates (Supplementary Fig. 2). To determine the incidence of AlMV in a greenhouse, 30 alstroemeria samples were collected and tested by RT-PCR. Results showed that 23 samples were positive for AlMV by PCR-gel electrophoresis and Sanger sequencing, suggesting a high incidence of AlMV infection. To the best of our knowledge, this is the first report of natural infection with AlMV in Alstroemeria in Korea. Further surveys of AlMV infection in greenhouses will help us prevent the spread of this viral disease in Alstroemeria.

First report of natural infection by two potyviruses on amaryllis (Hippeastrum hybridum) plants from India.

Amaryllis plants showing the chlorotic stripes and yellowing of the leaves were collected. Electron microscopy of the infected plants revealed flexuous filamentous particles and disease incidencewas recorded up to 64%. The presence of virus was confirmed using potyvirus genus and species-specific primers using Reverse transcription PCR. Blastn analysis showed that the samples were infected with leek yellow stripe virus (LYSV) and onion yellow dwarf virus (OYDV)the of genus Potyvirus, family Potyviridae. To the best of our knowledge, this is the first report of an association between LYSV (MZ203474 to MZ203478) and OYDV (MZ203479-MZ203482) with amaryllis. The result also showed that infected Amaryllis plants may act as a reservoir of LYSV and OYDV for economically important Allium crops.

First report of natural infection by Trypanosoma cruzi in secretions of the scent glands and myocardium of Philander opossum (Marsupialia: Didelphidae): Parasitological and clinicopathological findings.

Trypanosoma cruzi is the etiologic agent of American trypanosomiasis and can infect humans and different species of domestic and wild animals. The marsupials are important wild reservoirs of T. cruzi, aiding in the maintenance of this agent in sylvatic and peri-domestic environments. The objective of this study was to report the parasitological and clinicopathological findings of a natural infection by T. cruzi in one specimen of Philander opossum that originated from the Brazilian Amazon. The animal was captured in a forest fragment near a rural community with reports of human Chagas disease. T. cruzi infection was diagnosed by blood smear examinations, blood culture, scent glands secretion culture, histopathological examination, and nested-PCR. Positive samples were subjected to PCR to characterize the discrete typing units (DTUs) of T. cruzi. Characteristic trypomastigotes of T. cruzi were observed in the blood smear, and spheromastigotes, epimastigotes, and trypomastigotes were visualized in the cultures. Non-suppurative myocarditis associated with amastigote clusters was the principal histopathological finding. DNA from T. cruzi was detected in samples of blood, blood cultures, scent glands secretion cultures, cardiac muscles, and the spleen. The TcI and the TcII/V/VI group DTUs were detected in blood culture and scent glands secretion cultures. Infection by T. cruzi can cause myocarditis in P. opossum and DTUs TcI and TcII/V/VI group mixed infection can be detected in the acute phase. P. opossum can be a source of infection for triatomine vectors and has the potential source for direct transmission of T. cruzi by secretions from the scent glands. These data are important to improve the understanding of the complex enzootic transmission cycle of T. cruzi in the Brazilian Amazon.

First report of Stemphylium lycopersici causing leaf spot on hot pepper in China

Hot pepper is an exceedingly popular vegetable crop in China and is cultivated on 737 hectares in Beijing. A severe leaf disease with typical symptoms of spots with grey centres and dark brown borders was observed on hot pepper plants in Beijing in the winter of 2016. More than 50% of all plants in the region were infected. Fungal cultures were isolated from naturally infected leaf tissue, and identified as Stemphylium lycopersici based on morphological features, cultural characteristics and molecular identification by sequencing the ITS, gpd and cmdA genes. Pathogenicity was determined by inoculating healthy pepper plants with hyphal suspensions, and the fungus was re-isolated from developing lesions on the inoculated plants, thus fulfilling Koch’s postulates. This is the first report of natural infection by S. lycopersici causing leaf spot on hot pepper in China.

Nyssorhynchus dunhami: bionomics and natural infection by Plasmodium falciparum and P. vivax in the Peruvian Amazon.

BACKGROUND Nyssorhynchus dunhami, a member of the Nuneztovari Complex, has been collected in Brazil, Colombia, and Peru and described as zoophilic. Although to date Ny. dunhami has not been documented to be naturally infected by Plasmodium, it is frequently misidentified as other Oswaldoi subgroup species that are local or regional malaria vectors. OBJECTIVES The current study seeks to verify the morphological identification of Nuneztovari Complex species collected in the peri-Iquitos region of Amazonian Peru, to determine their Plasmodium infection status, and to describe ecological characteristics of their larval habitats. METHODS We collected Ny. nuneztovari s.l. adults in 2011-2012, and Ny. nuneztovari s.l. larvae and adults in 2016-2017. When possible, samples were identified molecularly using cytochrome c oxidase subunit I (COI) barcode sequencing. Adult Ny. nuneztovari s.l. from 2011-2012 were tested for Plasmodium using real-time PCR. Environmental characteristics associated with Ny. nuneztovari s.l. larvae-positive water bodies were evaluated. FINDINGS We collected 590 Ny. nuneztovari s.l. adults and 116 larvae from eight villages in peri-Iquitos. Of these, 191 adults and 111 larvae were identified by COI sequencing; all were Ny. dunhami. Three Ny. dunhami were infected with P. falciparum, and one with P. vivax, all collected from one village on one night. Ny. dunhami larvae were collected from natural and artificial water bodies, and their presence was positively associated with other Anophelinae larvae and amphibians, and negatively associated with people living within 250m. MAIN CONCLUSIONS Of Nuneztovari Complex species, we identified only Ny. dunhami across multiple years in eight peri-Iquitos localities. This study is, to our knowledge, the first report of natural infection of molecularly identified Ny. dunhami with Plasmodium. We advocate the use of molecular identification methods in this region to monitor Ny. dunhami and other putative secondary malaria vectors to more precisely evaluate their importance in malaria transmission.

Trypanosoma cruzi infection in naturally infected dogs from an endemic region of Cundinamarca, Colombia.

The seropositivity and risk factors for Trypanosoma cruzi infection in dogs from a municipality of Cundinamarca, a central state of Colombia were studied. A total of 356 client-owned dogs from urban, peri-urban and rural areas of La Mesa municipality, (Cundinamarca, Colombia) were randomly selected. Blood samples were collected by venipuncture. Anti-T. cruzi antibodies were determined using the enzyme-linked immunosorbent assay (ELISA) method. Reactive ELISA sera were processed by indirect immunofluorescence to confirm the presence of anti-T. cruzi antibodies. Chi-square tests were conducted for statistical analysis. Serologic tests for T. cruzi infection showed a prevalence of 29.49% (105/356), the rural area show a highest T. cruzi infection pattern in comparison with the other zone locations. Two triatomine species were found through the study: Panstrongylus geniculatus (53.4%) and Rhodnius colombiensis (46.6%). The prevalence of positive vectors for parasite was of 52.1% (38/73). Additionally, a very close relation between triatomine bugs and dogs in the rural zone (1:3.1) was observed. These results are the first report of natural infection by T. cruzi in domestic dogs in La Mesa municipality. In conclusion, the presence of anti-T. cruzi antibodies in dogs in this area suggest vector transmission. There is a need for active surveillance programs throughout the La Mesa municipality and vector control strategies should also be implemented.

Molecular detection of Histoplasma capsulatum in insectivorous and frugivorous bats in Southeastern Brazil.

Bats are considered to play a significant role in the epidemiology of histoplasmosis, worldwide. We investigated the occurrence of H. capsulatum in lung samples from 89 bats, from urban areas in Southeastern Brazil, using nested PCR based on ribosomal DNA. Fungal DNA was detected in 31/89 samples (34.8%), of which 13/31 were Molossids (41.9%), 4/31 Eumops spp. (12.9%), 2/31 Artibeus lituratus (6.5%), and 12/31 others (38.7%). This is the first report of natural infection by H. capsulatum in A. lituratus in Southeastern Brazil, which reinforces the importance of these synanthropic animals in the epidemiology of histoplasmosis in urban areas.

Assessment of the role of small mammals in the transmission cycle of tegumentary leishmaniasis and first report of natural infection with Leishmania braziliensis in two sigmodontines in northeastern Argentina.

To contribute to the knowledge of the role of small mammals in the transmission cycle of tegumentary leishmaniasis caused by Leishmania braziliensis, we studied the small mammal community and its temporal and spatial association with phlebotominae, as well as small mammal infection by Leishmania spp. by PCR-RFLP analyses in an endemic area of northeastern Argentina. Ten small mammal samplings were conducted (2007-2009, 7506 Sherman trap nights and 422 cage trap nights). In two of these samplings, 16 capture stations each one consisting of a CDC light trap to capture phlebotominae, two to four Sherman traps and two cage traps were placed. We found co-occurrence of phlebotominae and small mammal captures in four stations, which were all the stations with small mammal captures and yielded 97% (2295 specimens, including 21 gravid females) of the total phlebotominae captures, suggesting that small mammals may provide a potential source of blood for phlebotominae females. One Didelphis albiventris and two Rattus rattus were associated with high captures of Nyssomyia whitmani, vector of L. braziliensis in the study area. The PCR-RFLP analyses confirm the presence of L. braziliensis in two sigmodontine small mammals (Akodon sp. and Euryoryzomys russatus) for the first time in Argentina, to our knowledge.

First detection of natural infection of Aedes aegypti with Zika virus in Brazil and throughout South America

Zika virus (ZIKV) has caused a major epidemic in Brazil and several other American countries. ZIKV is an arbovirus whose natural vectors during epidemics have been poorly determined. In this study, 1,683 mosquitoes collected in the vicinity of ZIKV suspected cases in Rio de Janeiro, Brazil, from June 2015 to May 2016 were screened for natural infection by using molecular methods. Three pools of Aedes aegypti were found with the ZIKV genome, one of which had only one male. This finding supports the occurrence of vertical and/or venereal transmission of ZIKV in Ae. aegypti in nature. None of the examined Ae. albopictus and Culex quinquefasciatus was positive. This is the first report of natural infection by ZIKV in mosquitoes in Brazil and other South American countries. So far, Ae. aegypti is the only confirmed vector of ZIKV during the ongoing Pan-American epidemics.

First report of Stemphylium solani causing leaf spot on wild eggplant in China

Wild eggplant (Solanum torvum) has been used as a rootstock for eggplant propagation in China for decades. In the winter of 2011–2012, a severe disease with typical symptoms of small brown spots was observed on the leaves of wild eggplant in Chengmai County, Hainan Province, China. This disease had an incidence of about 30–65% among all the plants in the region (about 2 hectares in total). The causal pathogen was identified as Stemphylium solani based on morphological features, cultural characteristics and molecular identification. The pathogenicity was confirmed by Koch’s postulates. This is the first report of natural infection by S. solani causing leaf spot on wild eggplant in China.

FIRST REPORT OF A 16SrIX GROUP (PIGEON PEA WITCHES'-BROOM) PHYTOPLASMA ASSOCIATED WITH GRAPEVINE YELLOWS IN IRAN

Association of 16SrXII-A phytoplasmas with grapevine yellows (GY) has been previously reported from Fars and Lorestan provinces of Iran (Salehi et al., 2014). During 2013-2014 surveys, symptoms resembling those of GY disease including thickening and downward rolling of leaves, vein chlorosis and necrosis, shoot dieback and stunting were observed in vineyards of Shiraz area (Fars province, Iran). Total DNA samples extracted from eight symptomatic and eight symptomless vines were tested for phytoplasma presence. In direct PCR using primers P1/P7 and nested PCR using primer pairs P1/P7 followed by R216F2n/R16R2 (Lee et al., 1998), respectively amplicons of ca. 1.8 and 1.2 kb were amplified from symptomatic but not from symptomless vines. Samples that yielded P1/P7 amplicons (4/8) were cloned and sequenced; obtained sequences showed 100% identity. A BLAST search using full length 16S rRNA gene sequence of a representative of these sequences (GenBank Accession No. KX011516) revealed that the sequence shared 99% homology with Indian Brassica rapa phyllody phytoplasma (HM988986), a representative of 16SrIX-C subgroup. Computer-simulated restriction analysis using iPhyClassifier showed that the RFLP profile of the Shiraz GY (SGY) phytoplasma 16S rDNA sequence was identical (similarity coefficient 1.00) to Picris echioides yellows phytoplasmas (Y16389) representative of subgroup C of the 16SrIX group. Phylogenetic analysis revealed that SGY phytoplasma clustered with16SrIX group phytoplasmas closer to Indian B. rapa phyllody phytoplasma, a 16SrIX-C member. To our knowledge, this is the first report of natural infection of grapevine by a 16SrIX phtoplasma in Iran.

Molecular detection of Wolbachia pipientis in natural populations of mosquito vectors of Dirofilaria immitis from continental Portugal: first detection in Culex theileri.

Wolbachia pipientis (Rickettsiales: Rickettsiaceae) protects mosquitoes from infections with arboviruses and parasites. However, the effect of its co-infection on vector competence for Dirofilaria immitis (Spirurida: Onchocercidae) in the wild has not been investigated. This study aimed to screen vectors of D. immitis for wPip, to characterize these, and to investigate a possible association between the occurrence of W. pipientis and that of the nematode. The presence of W. pipientis was assessed in the five mosquito potential vectors of D. immitis in Portugal. Polymerase chain reaction (PCR) products were sequenced, and wPip haplotypes were determined by PCR-restricted fragment length polymorphism (RFLP). Results showed that wPip was detected in 61.5% of Culex pipiens (Diptera: Culicidae) pools and 6.3% of Culex theileri pools. wPip 16s rRNA sequences found in Cx. theileri exactly match those from Cx. pipiens, confirming a mosquito origin, rather than a nematode origin, as some specimens were infected with D. immitis. Only wPip haplotype I was found. No association was found between the presence of wPip and D. immitis in mosquitoes and hence a role for this endosymbiont in influencing vectorial competence is yet to be identified. This study contributes to understanding of wPip distribution in mosquito populations and, to the best of the authors' knowledge, is the first report of natural infections by wPip in Cx. theileri.

FIRST REPORT OF MYROTHECIUM RORIDUM CAUSING ROUND SPOT ON DENDROBIUM CANDIDUM IN CHINA

Dendrobium candidum (family Orchidaceae) is grown as a medicinal plants in China. In July 2013, in some plastic greenhouses of Puer (Southwest Yunnan) round spots were observed on the leaves of D. candidum plants with 20-30% incidence. Initial symptoms included black to brown circular or subcircular lesions 0.5-2.0 cm in diameter, surrounded by water-soaked tissue and later by concentric rings, grey brown and dark green. Infected leaves turned yellow, wilted, and decayed. Symptomatic leaf tissues (4x5 mm fragments) were surface-sterilized and cultured onto potato dextrose agar (PDA) at 25°C in darkness. Developing colonies were white, floccose at the beginning, then produced sporodo- chia in dark greyish-green concentric rings bearing slimy spore masses. Conidia were smooth, hyaline, cylindrical with rounded ends, 4.9-8.9x1.4-2.8 μm in size. DNA was extracted from a fungal isolate denoted YNPE-SH9 using a DNA extraction kit (OMEGA, USA). The internal tran- scribed spacer (ITS) region and 5.8S rDNA gene were am- plified using primer pair of ITS1 and ITS4 (White et al., 1990). BLAST search of the obtained sequence (GenBank accession No. KM986033) revealed a 100% identity with two Myrothecium roridum strains (AJ301995, AJ302001). Leaves of 10 healthy greenhouse-grown D. candidum plants at the seedling stage were inoculated by injecting a conidial sus- pension of M. roridum (1.1×105 conidia per ml) and kept at 28°C, 80% relative humidity and 14-h photoperiod. Seed- lings inoculated with sterile distilled water served as con- trols. Symptoms like those seen in naturally infected plants were observed in all inoculated D. candidum seedlings seven days post inoculation while control plants remained symp- tomless. To our knowledge, this is the first report of natural infection by M. roridum on Dendrobium spp. in China.

First Report of Protechinostoma mucronisertulatum (Echinostomatidae) in a Sandhill Crane (Grus canadensis) from Saskatchewan, Canada

ABSTRACT: We report a new host record for Protechinostoma mucronisertulatum. These small trematode parasites were found in a debilitated, immature, male sandhill crane (Grus canadensis) during autumn migration from the Canadian prairies. Necropsy examination identified fibrinonecrotizing and ulcerative jejunitis with a focal perforation and associated local fibrinous coelomitis. Cross sections of P. mucronisertulatum were present within lesions, although their role in the pathogenesis of these lesions is undetermined. Prior reports of natural infections attributed to these flukes are rare and have been limited primarily to sora rails (Porzana carolina) from the central North American flyway. Specimens in the sandhill crane were morphologically consistent with the original description; we provide the first complete series of measurements from flukes derived from a natural infection.

Natural infection of phlebotomines (Diptera: Psychodidae) by Leishmania (Leishmania) amazonensis in an area of ecotourism in Central-Western Brazil.

BackgroundBonito municipality, known as an area of ecoturism, in Mato Grosso do Sul state, Brazil, is also a focus of visceral and cutaneous leishmaniases, with cases registered in both human and canine populations. This study sought to investigate natural infection by flagellate forms of Leishmania in phlebotomines of the urban area of Bonito.FindingsSand flies were collected fortnightly from October 2005 to July 2006 with modified automatic light traps installed in peridomiciles and animal shelters in the center and on the outskirts of the city. The females were dissected and their guts observed under an optical microscope. A total of 1977 specimens were captured, Lutzomyia longipalpis (88.4 %) and Bichromomyia flaviscutelata (3.0 %) being the most frequent species. Bi. flaviscutellata was found infected by flagellates that were identified as Leishmania (Leishmania) amazonensis by indirect immunofluorescence reaction, employing monoclonal antibodies and the biotin-avidin system. This is the first report of natural infection by L. amazonensis in Bi. flaviscutellata in a Brazilian urban area.ConclusionsAs Bi. flaviscutellata is only slightly attracted by humans, the transmission of L. amazonensis in the study area may have a zoonotic character; however, the sympatric occurrence of this parasite and Lu. longipalpis should be taken into consideration by the local health authorities since this sand fly has already been found with L. amazonensis DNA in a focus of canine visceral leishmaniasis in Bonito municipality.

First report of Myxobolus sp. infection in the skeletal muscle of Neotropical freshwater fish Piaractus mesopotamicus.

Although there are several reports of Myxobolus species infecting the somatic muscle of teleost fish, species of this genus have not been described parasitizing the muscle tissue of pacus, Piaractus mesopotamicus. This study presented the first report of natural infection by Myxobolus sp. in the skeletal muscle of pacus reared in intensive system. Twenty-one fish (±142.2 g; ±23.1 cm) were captured (April 2013) from an intensive fish farm in São Paulo State, southeast Brazil. Spores of Myxobolus sp. were contained within plasmodia and showed oval morphology with the apical portion slightly pyriform and two polar capsules pyriform retaining the same ratio to each other. In the histopathological study, the skeletal muscle do not present signs of inflammation. The integrity of myofibrils within the infected fibers showed some degree of degeneration, with partial loss of myofibrillar details and striations. Spores were found infecting the skeletal muscle of 18 fish (85.7%). Finally, the high prevalence of Myxobolus sp. infection in the skeletal muscle of P. mesopotamicus and the absence of macroscopic lesions in the muscle tissue indicate the necessity of more meticulous examinations during the health management in the rearing system and at slaughter of pacus to ensure best quality of meat.

Epidemiological study on sand flies in an endemic focus of cutaneous leishmaniasis, bushehr city, southwestern iran.

Cutaneous leishmaniasis is the most important health problem in the city of Bushehr, southwestern Iran. The objective of the study was to determine some ecological aspects of sand flies in the city during 2010–2011. Sand flies were collected monthly from outdoors and indoors by sticky traps at four selected districts of the city. They were also dissected and examined by nested-PCR for identification of the parasite during August–September of 2011. A total of 1234 adult sand flies were collected and 6 species including 3 of Genus Phlebotomus and 3 of Genus Sergentomyia were identified. Four species including P. papatasi (3.98%), P. sergenti (1.14%), S. tiberiadis (87.18%), and S. baghdadis (7.7%) were found indoors. Six species including P. papatasi (3.47%), P. sergenti (3.17%), P. alexandri (0.1%), S. tiberiadis (77.74%), S. baghdadis (15.41%), and one female of S. clydei (0.11%) were collected from outdoors. Sand flies started to appear from March and disappear at the end of January. There was only one peak in the density curve in July. The study revealed that S. tiberiadis and S. baghdadis could enter indoors which 89 and 81.8% of them were found blood-fed, respectively. Moreover, P. papatasi, S. tiberiadis, and S. baghdadis were active indoors and outdoors in most months of the year. Nested-PCR of P. papatasi females was positive against kinetoplast DNA of L. major and L. turanica and also mixed natural infections were found by L. gerbilli and L. turanica. Moreover, mixed infections by L. major and L. turanica were observed in this species. Sergentomyia clydei and S. tiberiadis were found to be negative to any DNA of Leishmania species. Phlebotomus sergenti females were found infected with DNA of L. turanica and this is the first report of natural infection and detection of the parasite from this sand fly species in worldwide.

First report of natural infection in hedgehogs with Leishmania major, a possible reservoir of zoonotic cutaneous leishmaniasis in Algeria

We report here the first known cases of natural infection of hedgehogs with Leishmania major. Cutaneous leishmaniasis is an important public health problem in the area of M'sila, a semi-arid province in Algeria's northern Sahara, where two species of hedgehog live, Atelerix algirus and Paraechinus aethiopicus. The aim of this research was to survey Leishmania infection in these hedgehogs and evaluate whether they were reservoir hosts of Leishmania in an endemic zoonotic focus of leishmaniasis. Serological and molecular methods were used to determine the presence of Leishmania in 24 hedgehogs caught directly by hand and identified at species level as 19 A. algirus and 5 P. aethiopicus. Specific anti-Leishmania antibodies were detected in 29.2% of individuals by Western blot and in 26.3% by ELISA. The real-time PCR performed in spleen, ear and blood samples detected Leishmania spp. DNA in 12.5% of the individuals, one A. algirus and two P. aethiopicus. Three skin and two spleen samples of these animals were found to be parasitized and were identified by molecular test as L. major. Considering our results, it is suggested that hedgehogs have a potential epidemiological role as reservoir hosts of L. major.

Neotropical echinococcosis: Second report of Echinococcus vogeli natural infection in its main definitive host, the bush dog (Speothos venaticus)

The bush dog (Speothos venaticus) acts as the natural definitive host in the life cycle of Echinococcus vogeli, the causative agent of polycystic hydatid disease, a zoonotic neglected disease in the South America. We report a case of natural infection by Echinococcus vogeli in a bush dog from the Brazilian Amazon, confirmed by the morphological and morphometric examination of adult parasites and their hooks obtained from the small intestine of the canid. Additionally, mitochondrial DNA sequence analysis corroborated these findings. This is the second report of natural infection by E. vogeli in a bush dog.