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Background: The current study aimed to determine the prevalence of cutaneous leishmaniasis (CL) in Al-Muthanna province (Iraq) and to characterize the Leishmania species that cause cutaneous lesions through conventional polymerase chain reaction techniques in some patients during the first 7 months of the year 2020., Materials and Methods: Medical information on patients with CL was obtained from archived records at the Al-Muthanna Health Office's Public Health Department (2015-2020). In the Al-Hussein Teaching Hospital laboratory, 95 CL samples were collected and examined microscopically for molecular characterization using Giemsa staining., Results: Between 2015 and 2020, 2,325 patients (1,184 men and 1,141 women) were enrolled. Although CL occurred across all age groups, those aged range of 5-14 years had the highest proportion of infections (53.0%). This study found that most infections occurred between December and February, peaking in January. Only 63 of 95 CL samples were positive for the Internal Transcribed Spacer 1 region. L. tropica was found in 39 samples (61.9%), whereas L. major was found in 24 samples (38.1%), in CL patients. Although dermal lesions develop in all body regions, a single lesion is the most common. The upper limbs (13 of 16 samples, 33.3%)were infected with L. tropica , whereas the lower limbs (9 of 14 samples, 37.5%) were infected with L. major . In contrast to L. major , most L. tropica lesions occur in urban areas., Conclusion: Our study indicates that CL is endemic in the Al-Muthanna province and that two Leishmania spp. coexist in the province. Molecular diagnosis is a vital component in determining many clinical symptoms of the Leishmania parasite as well as implementing suitable therapeutic, epidemiological, and control strategies., Competing Interests: No conflict of interest., (© 2024 by The Korean Society of Infectious Diseases, Korean Society for Antimicrobial Therapy, The Korean Society for AIDS, and Korean Society of Pediatric Infectious Diseases.)

Objective: Leishmaniasis is a parasitic disease caused by more than 20 Leishmania species. This disease is spread by vectors. Many researchers agree that Leishmania was spread to mammals by sandflies of the genus Phlebotomus and Lutzomyia. Leishmaniasis is still considered one of the most neglected diseases by the World Health Organization (WHO). An estimated 0.7-1 million new cases of leishmaniasis are reported annually from approximately 100 endemic countries. The types of leishmaniasis in humans are the visceral (VL), cutaneous (CL), mucocutaneous (MCL), diffuse cutaneous (DCL), and post kala-azar dermal (PKDL) forms of Leishmaniasis. The aim of this study is to perform phylogenetic analysis of Leishmania origin based on ITS1 gene region using insilico techniques. In this way, it is also aimed to take a snapshot of a meta-analysis of vertical and horizontal spread at the global level. Methods: In this study, Leishmania ITS1 region sequences presented with the GenBank data of the National Center for Biotechnology Information, USA, (NCBI) until 15.05.2019 were taken and analyzed by in-silico techniques. 914 sequences were obtained for the Leishmania ITS1 region in the NCBI database. All sequences were examined and sequences without indel problems were selected from these strains mapped according to the consensus sequence. It was decided to form a phylogenetic tree with the forms that were examined and 65 strains were obtained by removing the sub-branches. Results: The phylogenetic tree obtained in this study showed that Leishmania strains clustered in six branches according to the ITS1 region. Here, a phylogenetic tree is drawn and the molecular epidemiological and demographic data of these six generations and beyond, which are obtained as a result of the genetic relationships between the strains, are summarized. Conclusion: In conclusion, Leishmaniasis is an important public health problem that can be seen in many developing countries. In this study, the strains examined using the in-silico method were isolated from different geographies of the world between 1984 and 2018. The phylogenetic relationships between these strains show not only the vertical spread of the origins over the years, but also the horizontal spread geographically. These species were obtained from different host and tissue types. Thus, the relationships of Leishmania strains in the host-vector-reservoir chain are explained. Therefore, it is clear that there is a need for more meta-analysis studies such as this study on factors and their diffusion. [ABSTRACT FROM AUTHOR]

This is the first study on phylogenetic relationships in the genus Artemia Leach, 1819 using the pattern and sequence of secondary structures of internal transcribed spacer 1 (ITS1). Significant intraspecific variation in the secondary structure of ITS1 rRNA was found in Artemia tibetiana. In the phylogenetic tree based on joined primary and secondary structure sequences, Artemia urmiana and parthenogenetic populations displayed new lineages, and two New World species (Artemia franciscana and Artemia persimilis) were located in a basal clade that was not detected in previous studies. The close evolutionary relationship between A. franciscana and A. persimilis are expressively supported by the previous empirical and experimental investigation on the ability of hybridization (in natural habitats and lab conditions) and analysis on allozyme markers.

Angiostrongylus cantonensis is a parasitic nematode known to infect humans through the ingestion of third stage larvae which can cause inflammation and damage to the central nervous system. Currently, polymerase chain reaction (PCR) is one of the most reliable diagnostic methods for detecting A. cantonensis in humans as well as in gastropod hosts, but requires expensive and specialized equipment. Here, we compare the sensitivity and accuracy of a recombinase polymerase amplification Exo (RPA-EXO) assay, and a recombinase polymerase amplification lateral flow assay (RPA-LFA) with a traditional quantitative PCR (qPCR) assay currently available. The three assays were used to test 35 slugs from Hawai'i for the presence of A. cantonensis DNA. Consistent results among the three tests were shown in 23/35 samples (65.7%), while 7/35 (20%) were discordant in low infection level samples (<0.01 larvae per mg tissue), and 5/35 (14.3%) were equivocal. To evaluate sensitivity, a partial ITS1 gene was cloned, and serial plasmid dilutions were created ranging from 100 copies μL-1 to ~1 copy μL-1. All three assays consistently detected 50-100 copies μL-1 in triplicate and qPCR was able to detect ~13 copies μL-1 in triplicate. RPA-EXO was able to detect 25 copies μL-1 in triplicate and RPA-LFA was not able to amplify consistently below 50 copies μL-1. Thus, our RPA-EXO and RPA-LFA assays do not appear as sensitive as the current qPCR assay at low DNA concentrations; however, these tests have numerous advantages that may make them useful alternatives to qPCR.

Background: Leishmaniasis is endemic in Iran. Different species of Leishmania (L.) parasites are causative agents of this disease. Correct identification of Leishmania species is important for clinical studies,prevention, and control of the diseases. Mix up of Leishmania isolates is possible in the laboratory, so there is need for verification of species for isolates of uncertain identity. Different methods may be used for this purpose including isoenzyme electrophoresis and molecular methods. The isoenzyme lectrophoresis, due to its drawbacks, is feasible only in specialized laboratories while molecular methods may be more feasible. The aim of this research was to study the application of the internal transcribedspacer 1 (ITS1) sequencing method, in comparison to isoenzyme electrophoresis method, for verification of Leishmania species.Methods: Six Leishmania isolates were received from different research institutions in Iran. The species of these isolates were known by donating institution according to their isoenzyme profile. The species of these isolates were re-identified in Pasteur Institute of Iran by PCR amplification of ITS1 followed bysequencing and comparison of these sequences with Leishmania sequences in GenBank. Isoenzyme electrophoresis was performed for confirmation of the results of ITS1.Results: ITS1 sequence showed that some isolates were mixed up or contaminated with Crithidia. Isoenzyme electrophoresis confirmed the results of ITS1 sequences.Conclusion: ITS1 sequencing is relatively more feasible than the traditional isoenzyme electrophoresismethod and is suggested for verification of Leishmania species.

Background: Trichomoniasis is a worldwide protozoan parasitic disease and metronidazole is a choice drug for its treatment. Because of disease importance in public health and its controversial ideas about the prevalence of drug resistance, this study was carried out.Methods: Fifty-two suspected vaginal samples were collected from 2006 to 2007 in Gynecology Maryam Hospital, Tehran, Iran. All isolates were examined by microscopic, culture and PCR tech­niques. The PCR products were analyzed by RFLP and CSGE methods and two suspected samples were sequenced.Results: Trichomonas vaginalis was identified from all 52 samples. Of 52 isolates, 45 samples were successfully cultured and amplified by PCR except one. Seven were positive only by PCR. Finally, ITS1 fragment was successfully amplified in 51 of 52. CSGE analysis and PCR products di­gestion by MspI followed by sequencing showed nucleotide mutation at position 209 (C209T) of the ITS1 fragment in two (3.9%) of them.Conclusion: The results showed mutation in ITS1 fragment of T. vaginalis in two (3.9%) of Ira­nian isolates which may be related to metronidazole resistance.

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Background: Leishmaniasis is endemic in Iran. Different species of Leishmania (L.) parasites are causative agents of this disease. Correct identification of Leishmania species is important for clinical studies, prevention, and control of the diseases. Mix up of Leishmania isolates is possible in the laboratory, so there is need for verification of species for isolates of uncertain identity. Different methods may be used for this purpose including isoenzyme electrophoresis and molecular methods. The isoenzyme electrophoresis, due to its drawbacks, is feasible only in specialized laboratories while molecular methods may be more feasible. The aim of this research was to study the application of the internal transcribed spacer 1 (ITS1) sequencing method, in comparison to isoenzyme electrophoresis method, for verification of Leishmania species. Methods: Six Leishmania isolates were received from different research institutions in Iran. The species of these isolates were known by donating institution according to their isoenzyme profile. The species of these isolates were re-identified in Pasteur Institute of Iran by PCR amplification of ITS1 followed by sequencing and comparison of these sequences with Leishmania sequences in GenBank. Isoenzyme electrophoresis was performed for confirmation of the results of ITS1. Results: ITS1 sequence showed that some isolates were mixed up or contaminated with Crithidia. Isoenzyme electrophoresis confirmed the results of ITS1 sequences. Conclusion: ITS1 sequencing is relatively more feasible than thetraditional isoenzyme electrophoresis method and is suggested for verification of Leishmania specie.

Background: Cutaneous leishmaniasis is a vector-borne disease transmitted by biting of the sandfly, it is a severe health problem in many countries and endemic in most regions of Iraq. Objectives: This study was conducted to find the best method for diagnosis of cutaneous leishmaniasis, detect the genotypes of Leishmania tropica and Leishmania major in Ramadi (Iraq) by PCR-RFLP technique. Materials &methods: One hundred twenty-two patients 68 were males while the females gender were 54 with age ranged 1-68 years, CL who attended to Department of Dermatology in Al-Ramadi Teaching Hospital and dermatology Private clinics, during the period between November 2017 to April 2018. The Molecular study was carried out to detect the ITS1 gene by (PCR). The restriction fragment length polymorphism (RFLP) was adopted on ITS1-PCR product and after HaeIII digestion at 37C° for 2 hours. Results: Laboratory examination of 122 cases showed 62 infection cases in Cutaneous leishmaniasis by using PCR technique and in infection proportion reaches at 51% out of the total number of the cutaneous cases which are similar to leishmaniasis during the months of the study. The demographic study dealt with age, gender, number of lesions and body site of infection, demonstrated that the majority of patients at the age of 1-10 years with percent 28.7%. Also Males (55.7%) had higher infection than females (44.3%), upper limbs had the highest percentage (48%) when compared with other sites of infection, single lesion was documented in 55% of patients, while two lesions were observed in 25% and multiple (3-10) lesions were observed in 20%. Different techniques were used for diagnosis of CL including routine method performed by direct microscopic smear from lesion which showed amastigotes in the macrophage in 50 (41%) positive case. The Molecular study was carried out to detect the ITS1 gene (internal transcribed spacer1) by (PCR). DNA extracted from 122 samples showed 62 (51%) were positive for (ITS1)gene, The restriction fragment length polymorphism (RFLP) was adopted on ITS1-PCR product and after HaeIII digestion at 37Cº for 2 hours obtained two fragments of 60 and 200 bp 42 as L.tropica, and two fragments of 140 and 210 bp were identified 20 as L.major, genotype techniques were performed for all positive samples. Conclusion: CL is highly spread with single lesions more than multiple lesions and molecular detection showed that L.tropica more common than L.major. [ABSTRACT FROM AUTHOR]

In Thailand, there are currently five recognized species members of the bubble-nesting Betta genus, namely Betta splendens, B. smaragdina, B. imbellis, B. mahachaiensis and B. siamorientalis. In 2010, we indicated the possibility, based on COI barcoding evidence, that there might be two additional species, albeit cryptic, related to the type-locality B. smaragdina in some provinces in the northeast of Thailand. In the present study, after a more extensive survey of the northeast, and phylogenetic analyses based on COI and ITS1 sequences, the B. smaragdina group may be composed of at least 3 cryptic species members. The phylogenetic positions of these B. smaragdina group members in the bubble-nesting bettas' tree together with those of their congeners have been consolidated by better DNA sequence quality and phylogenetic analyses. With a better supported tree, the species statuses of B. siamorientalis and the Cambodian B. smaragdina-like fish, B. stiktos, are also confirmed.

Background: Leishmaniasis is endemic in Iran. Different species of Leishmania (L.) parasites are causative agents of this disease. Correct identification of Leishmania species is important for clinical studies, prevention, and control of the diseases. Mix up of Leishmania isolates is possible in the laboratory, so there is need for verification of species for isolates of uncertain identity. Different methods may be used for this purpose including isoenzyme electrophoresis and molecular methods. The isoenzyme electrophoresis, due to its drawbacks, is feasible only in specialized laboratories while molecular methods may be more feasible. The aim of this research was to study the application of the internal transcribed spacer 1 (ITS1) sequencing method, in comparison to isoenzyme electrophoresis method, for verification of Leishmania species., Methods: Six Leishmania isolates were received from different research institutions in Iran. The species of these isolates were known by donating institution according to their isoenzyme profile. The species of these isolates were re-identified in Pasteur Institute of Iran by PCR amplification of ITS1 followed by sequencing and comparison of these sequences with Leishmania sequences in GenBank. Isoenzyme electrophoresis was performed for confirmation of the results of ITS1., Results: ITS1 sequence showed that some isolates were mixed up or contaminated with Crithidia. Isoenzyme electrophoresis confirmed the results of ITS1 sequences., Conclusion: ITS1 sequencing is relatively more feasible than the traditional isoenzyme electrophoresis method and is suggested for verification of Leishmania species.

Background: Trichomoniasis is a worldwide protozoan parasitic disease and metronidazole is a choice drug for its treatment. Because of disease importance in public health and its controversial ideas about the prevalence of drug resistance, this study was carried out., Methods: Fifty-two suspected vaginal samples were collected from 2006 to 2007 in Gynecology Maryam Hospital, Tehran, Iran. All isolates were examined by microscopic, culture and PCR techniques. The PCR products were analyzed by RFLP and CSGE methods and two suspected samples were sequenced., Results: Trichomonas vaginalis was identified from all 52 samples. Of 52 isolates, 45 samples were successfully cultured and amplified by PCR except one. Seven were positive only by PCR. Finally, ITS1 fragment was successfully amplified in 51 of 52. CSGE analysis and PCR products digestion by MspI followed by sequencing showed nucleotide mutation at position 209 (C209T) of the ITS1 fragment in two (3.9%) of them., Conclusion: The results showed mutation in ITS1 fragment of T. vaginalis in two (3.9%) of Iranian isolates which may be related to metronidazole resistance.

Extremely long PCR fragments were generated by PCR amplification of ITS and 5.8S rDNA from Cochlodinium polykrikoides against other dinoflagellates. These patterns were consistent among geographically different isolates of C. polykrikoies. DNA sequencing reactions revealed that the PCR products were 1,166 bp in length and consisted of 813 bp of ITS1, 160 bp of 5.8S rDNA and 193 bp of ITS2. Thus, the long length was caused mainly by the long ITS1 sequence. Cryptically, the ITS1 contained a tract of 101 bp that occurs six times in tandem. The six repeated elements had identical nucleotide sequences. ITS1, therefore, separated three distinct regions: the 5' end (122 bp), the six parallel repeats (606 bp), and the 3' region (85 bp). Interestingly, both the single and six-repeat sequences should be palindrome-like sequences. In inferred secondary structures, both repeat sequences formed a long helical structure. This is the first reported discovery of comparatively long internal repeats in the ITS1 of dinoflagellates.

Background: The Leishmaniasis is a zoonosis disease with a global spread that occurs in three, cutaneous, mucocutaneous and visceral forms. In Iran, three species of Leishmania tropica, Leishmania major, and Leishmania infantum have been reported from different regions of the country. Nowadays, molecular methods are usually used for the diagnosis and identification of parasite species. Amplification of the ITS1 gene can also be used to differentiate symptomatic and asymptomatic visceral leishmaniasis as well as types of cutaneous leishmaniasis. Objectives: The main aim of our research was to diagnose and identify the common Leishmania species in Iran through ITS1 gene amplification and using the PCR-RFLP method. Methods: First, L. major, L. tropica, and L. infantum parasites were proliferated in an RPMI culture medium, and DNA was extracted from these species separately then ITS1 gene of the parasite was amplified using the PCR-RFLP method. Results: The result of PCR-RFLP after enzymatic cutting indicated, L. tropica produced 4 fragments of 139, 76, 56, 20 bp bands; L. major showed two fragments of 165, 139 bp bands and L. infantum produced three fragments of 141, 91, 54 bp bands. Conclusion: The results of the present study showed that the use of ITS1 gene and HaeIII enzyme in the PCR-RFLP method is efficient for identifying L. tropica, L. major, and L. infantum. [ABSTRACT FROM AUTHOR]

Background/Objective: The cestode Echinococcus granulosus causes cystic echinococcosis, a zoonotic parasitic infection that constitutes a significant public health risk. This parasite has been documented to have potential reservoirs and carriers among wild canids, namely wolves, foxes and jackals. This study aimed to determine the prevalence and molecular characteristics of E. granulosus sensu lato species/genotypes among wild canids in three northern, northeastern and north‐western Iran regions. Methods: From 2019 to 2022, 93 wild canid carcasses (69 jackals), (22 foxes) and (2 wolves) were collected that were killed in car accidents or illnesses. Analyses of morphology and morphometry were performed to verify the presence of E. granulosus. To determine E. granulosus s.l. species/genotypes, polymerase chain reaction (PCR)‐RFLP (ITS1) was performed utilizing the Bsh1236I (BstUI) restriction enzyme. COX1, NADH1 and ITS1 gene sequencing were also performed to confirm the PCR‐RFLP results. Results: During this study, 93 wild canids were examined, and 3.2% (95% CI: 0%–7%) of the 93 were infected with Echinococcus. The north‐western region of Iran showed two out of 30 jackals (6.6%) infected with adult Echinococcus compared to one out of 35 jackals (2.8%) in the northern region. DNA from Echinococcus was detected in these individuals by PCR. Based on PCR‐RFLP analysis of the ITS1 gene and sequencing of COX1, NADH1 and ITS1 gene, E. granulosus sensu stricto genotype was confirmed in the jackals that had been infected. Conclusion: Evidence shows that E. granulosus occurs in jackals in Iran, with the E. granulosus s.s. genotype being the most common. This parasite has been identified as a zoonotic parasite with a genotype that can be transmitted to livestock and humans. Establishing effective control measures to prevent the spread of echinococcosis and ensure public health is crucial. [ABSTRACT FROM AUTHOR]

Comprehending the phylogeography of invasive organisms enhances our insight into their distribution dynamics, which is instrumental for the development of effective prevention and management strategies. In China, Pomacea canaliculata and Pomacea maculata are the two most widespread and damaging species of the non-native Pomacea spp.. Given this species’ rapid spread throughout country, it is urgent to investigate the genetic diversity and structure of its different geographic populations, a task undertaken in the current study using the COI and ITS1 mitochondrial and ribosomal DNA genes, respectively. The result of this study, based on a nationwide systematic survey, a collection of Pomacea spp., and the identification of cryptic species, showed that there is a degree of genetic diversity and differentiation in P. canaliculata, and that all of its variations are mainly due to differences between individuals within different geographical populations. Indeed, this species contains multiple haplotypes, but none of them form a systematic geographical population structure. Furthermore, the COI gene exhibits higher genetic diversity than the ITS1 gene. Our study further clarifies the invasive pathways and dispersal patterns of P. canaliculata in China to provide a theoretical basis. [ABSTRACT FROM AUTHOR]

Background: Leishmaniasis is a major medical health problem and distributes in nearly half of 31 provinces of Iran. We aimed to identify cutaneous and visceral Leishmania spp. isolated from infected humans and domestic dogs in various regions of Iran, 2010‒2013.Methods: DNA was extracted from 108 lesion exudate samples of suspected patients to cutaneous leishmaniasis and nine liver and spleen aspirates of infected dogs cultured in RPMI-1640 and amplified using partial sequence of ITS1 gene. The PCR amplicons were digested using HaeIII endonuclease enzyme and used in restriction fragment length polymorphism (RFLP) assay. Then, 48 amplicons representing various hosts were sequenced and compared to se­quences from GenBank databases using BLAST.Results: PCR-RFLP analysis showed that 60 and 48 CL patients were infected by Leishmania tropica and L. major, respectively. From nine canine visceral leishmaniasis (CVL) isolates, eight isolates were identified as L. infantum and one as L. tropica. The greatest similarity of 95.7% in ITS1 region was seen between L. infantum and L. major. Furthermore, the lowest similarity with 65.7% was seen between L. tropica and L. major. Intra-species comparison of ITS1 region in L. infantum, L. major and L. tropica isolates were showed 100%, 98.2% and 72.4 % similarities, respectively.Conclusion: PCR-RFLP based on ITS1 region is an appropriate method to distinguish three Leishmania spp. of L. major, L. tropica, and L. infantum. In intra-species comparison of ITS1 region, genotypic variations showed that L. tropica isolates were more heterogeneous than L. major and L. infantum isolates.

Humans can be infected with anthroponotic (Ancylostoma duodenale and Necator americanus) and with zoonotic (Ancylostoma ceylanicum, A. caninum, A. braziliense, and Uncinaria stenocephala) hookworms from dogs. Anthroponotic species are usually thought not to infect dogs. We used the internal transcribed spacer–1 (ITS1) gene in a quantitative PCR to detect anthroponotic and zoonotic hookworm species in fecal samples from 54 children and 79 dogs living in an indigenous community in tropical Northwestern Ecuador. Hookworm DNA was detected in 59.3% of children and 92.4% of dogs. Among samples from children, zoonotic hookworms were detected in 24.1% (A. ceylanicum 14.8%, A. caninum 11.1%, and A. braziliense 1.9%), whilst in dog samples, anthroponotic species were detected in 19.0% (N. americanus 12.4% and A. duodenale 6.3%). Sanger sequencing was performed successfully on 60 qPCR-positive samples (16 from children and 44 from dogs), and consensus sequences were obtained with >98% homology to GenBank references for hookworm spp. Phylogenetic analysis showed a close relationship between anthroponotic and zoonotic Ancylostoma species and no heterogeneity between A. duodenale and A. caninum; in human samples, we found A. ceylanicum but not A. braziliense sequences and we were unable to identify N. americanus in the dog samples. No infections with U. stenocephala were detected. Our data provide evidence for high rates of hookworm infections in indigenous children and dogs in a marginalized rural setting in coastal Ecuador. We also found evidence for potential cross-transmission of hookworm spp. between humans and dogs that represent a potential domestic reservoir for zoonotic and anthroponotic hookworms. [ABSTRACT FROM AUTHOR]

Cystic echinococcosis is an important cosmopolitan parasitic zoonosis that causes public health and economic problems in Egypt. The present study was undertaken to identify genotypes of hydatid cyst (HC) DNA isolated from different animal isolates and to identify the genotype of secondary hydatid cysts (HCs) developed in rabbits experimentally infected with camel HC for detection of any genetic mutation. In the present study, we extracted DNA from the germinal layers of 8 HCs collected from 3 camels, 1 cattle, 1 sheep and 3 donkeys in addition to 3 secondary HCs collected from rabbits experimentally infected with camel HC. PCR amplification of the ITS1 gene of all examined samples showed an amplified DNA band at 1115 bp. The partial nucleotide sequences of the ITS1 gene of all isolates were aligned and compared with the reference sequences of the genotypes G1–G8 in GenBank. The camel and rabbit samples were identified as Echinococcus canadensis genotype 6 (G6), while the cattle and sheep samples belonged to E. granulosus sensu stricto (G1). The donkey isolates belonged to E. equines (G4). Alignment of the ITS1 partial nucleotide sequences of the camel HCs and rabbit secondary HCs isolates with the G6 partial nucleotide sequence in GenBank was performed. Both camel HCs and rabbit secondary HCs isolates exhibited the same sequence identity matrix, which indicated the absence of mutation in the rabbit secondary HCs. It can be concluded that camel and rabbit samples were identified as E. canadensis (G6), the cattle and sheep samples belonged to E. granulosus sensu stricto (G1) and donkey isolates belonged to E. equines (G4). No mutation occurred during HCs transmission from camel to rabbit. [ABSTRACT FROM AUTHOR]

Neospora caninum is a protozoan coccidian parasite that can act as a cause of abortion in sheep. The aim of this study was to investigate the presence of this parasitic agent and its role in causing abortion in sheep of Iran. Between June 2019 and February 2022, 100 samples [brain (n = 39), placenta (n = 8), embryonic membrane (n = 7), cotyledon (n = 7), umbilical cord (n = 2), homogenate mixture of tissues (heart, liver, spleen and digestive track) (n = 37)] that were collected following the necropsies of 39 aborted ovine fetuses from different parts of the Alborz and Qazvin provinces, the north of the central region of Iran were employed for DNA extraction. Nc-5 was selected as the target gene sequence for amplification of DNA by using four pairs of primers in two semi-nested PCR. Samples considered positive for the presence of the NC-5 gene were examined to further confirm the presence of the ITS1 gene. Sequence of NC-5 gene was detected from the 27 tissue samples of 23 aborted ovine fetuses. The ITS1 gene sequence was detected in all of the 27 tissue samples that were positive for the NC-5 gene analysis. Brain tissue was the most studied tissue, and the highest number of positive cases was observed in this tissue. The present study updated the situation of ovine neosporosis in the central region of Iran and confirmed the presence of the N. caninum among sheep flocks' abortion. [ABSTRACT FROM AUTHOR]

The aim of this study was to characterize the Tunisian Fasciola spp. flukes by morphometric and molecular analyses. Flukes were collected from livers of sheep slaughtered in Sejnane slaughterhouses (Bizerte gouvernorate, Northwest Tunisia) between January and March 2021. Five morphometric parameters were determined for all the liver flukes, as follows: (i) total body length (BL), (ii) distance between ventral sucker and the tail (VS-T), (iii) distance between oral sucker and ventral sucker (OS-VS), (iv) abdomen diameter (AD), (v) tail diameter (TD) and the body length to width ratio (BL/BW). Molecular identification of the fluke specimens was carried out by polymerase chain reaction, restriction fragment polymorphism (PCR-RFLP) of a 680 bp sequence of the internal transcribes spacer 1 (ITS1) gene and by amplification, sequencing, and phylogenetic analysis of a 500 bp sequence of the ITS2 gene. Morphometric measurements showed that the mean of the total body length of the adult flukes was 21.1 ± 2.7 mm with minimum and maximum lengths of 13 and 31 mm, respectively. The PCR-RFLP analysis revealed a single profile consisting of three bands of approximately 370, 100, and 60 bp. Fasciola sequences described in the present study (GenBank numbers: OQ457027 and OQ457028) showed 99.58–100% identity to Fasciola hepatica. In conclusion, the results of this study show that molecular and phylogenetic analyses confirm the presence of a single species of F. hepatica in the Sejnane region Northwest of Tunisia. However, further studies are needed to identify the occurrence of Fasciola species in other Tunisian regions. [ABSTRACT FROM AUTHOR]

The rapid reduction in coral reefs worldwide has led to increasing attention toward protecting and restoring coral reef ecosystems. Coral reefs not only have a rich diversity of coral species, but they can also provide important products and services for human beings. One type of coral, Echinogorgia, has important scientific research value and application prospects. To understand the diversity of coral species, diving surveys were conducted in Xiamen Bay in 2017 and 2021, and a total of 928 samples were collected. Taxonomic research was conducted using methods such as morphological identification through electron microscopy. Specific phylogenetic trees of the COI gene, mtMuts gene, and ITS1 gene were analyzed. There were 47 specimens of Echinogorgia coral included among 928 samples. Fifteen species of Echinogorgia were identified, including Echinogorgia ramosa, Echinogorgia flexilis, Echinogorgia russelli, Echinogorgia ramulosa, and Echinogorgia gracilima (which represent the newly recorded species in the waters of China). This study increases the species diversity records in China and contributes to new geographical distribution information of Echinogorgia worldwide. The primary data also serve as the baseline data for long-term biomonitoring programs to estimate the status of octocorals in Xiamen Bay. [ABSTRACT FROM AUTHOR]

Background: Leishmaniasis is a major medical health problem and distributes in nearly half of 31 provinces of Iran. We aimed to identify cutaneous and visceral Leishmania spp. isolated from infected humans and domestic dogs in various regions of Iran, 2010-2013. Methods: DNA was extracted from 108 lesion exudate samples of suspected patients to cutaneous leishmaniasis and nine liver and spleen aspirates of infected dogs cultured in RPMI-1640 and amplified using partial sequence of ITS1 gene. The PCR amplicons were digested using HaeIII endonuclease enzyme and used in restriction fragment length polymorphism (RFLP) assay. Then, 48 amplicons representing various hosts were sequenced and compared to sequences from GenBank databases using BLAST. Results: PCR-RFLP analysis showed that 60 and 48 CL patients were infected by Leishmania tropica and L. major, respectively. From nine canine visceral leishmaniasis (CVL) isolates, eight isolates were identified as L. infantum and one as L. tropica. The greatest similarity of 95.7% in ITS1 region was seen between L. infantum and L. major. Furthermore, the lowest similarity with 65.7% was seen between L. tropica and L. major. Intra-species comparison of ITS1 region in L. infantum, L. major and L. tropica isolates were showed 100%, 98.2% and 72.4 % similarities, respectively. Conclusion: PCR-RFLP based on ITS1 region is an appropriate method to distinguish three Leishmania spp. of L. major, L. tropica, and L. infantum. In intra-species comparison of ITS1 region, genotypic variations showed that L. tropica isolates were more heterogeneous than L. major and L. infantum isolates. [ABSTRACT FROM AUTHOR]

Background: Cutaneous leishmaniasis is a disease transmitted by sand fly bites. This disease is highly prevalent in Syria where Leishmania major and Leishmania tropica are the known aetiological agents. In 2011, more than 58,000 cases were reported in the country by the Ministry of Health. The central region of the country harbors 20 % of the reported cases. However, the epidemiology of the disease in this area is not well understood. An epidemiological survey was conducted in 2010 to identity the circulating parasite and the sand fly vector in the central provinces of Edlib and Hama. Methods: Sand fly specimens were collected using CDC light traps and identified morphologically. Total DNA was extracted from the abdomens of female specimens and from Giemsa-stained skin lesion smears of 80 patients. Leishmania parasites were first identified by sequencing the ITS1 gene amplicons. Then polymorphism analysis was performed using the RFLP technique. Results: A total of 2142 sand flies were collected. They belonged to eight species, among which Phlebotomus sergenti and Phlebotomus papatasi were the most predominant. L. tropica ITS1 gene was amplified from two pools of P. sergenti specimens and from skin smears of cutaneous leishmaniasis patients. This suggests that P. sergenti is the potential vector species in the study area. The digestion profiles of the obtained amplicons by TaqI restriction enzyme were identical for all analysed L. tropica parasites. Moreover, L. infantum ITS1 gene was amplified from two pools of Phlebotomus tobbi in the relatively humid zone of Edlib. Conclusions: L. tropica is confirmed to be the aetiological agent of cutaneous leishmaniasis cases in the central provinces. RFLP technique failed to show any genetic heterogeneity in the ITS1 gene among the tested parasites. The molecular detection of this parasite in human skin smears and in P. sergenti supports the vector status of this species in the study area. The detection of L. infantum in P. tobbi specimens indicates a potential circulation of this parasite in the humid zone of Edlib. Further epidemiological studies are needed to evaluate the burden of this visceral parasite in the study region. [ABSTRACT FROM AUTHOR]