Cytb Gene Research Articles

N-Butylphthalide Potentiates the Effect of Fluconazole Against Drug-Resistant Candida glabrata and Candida tropicalis. Evidence for Its Mechanism of Action.

To define the antifungal activity of n-butylphthalide alone or in combination with fluconazole in Candida glabrata and Candida tropicalis. The antifungal activity of n-butylphthalide alone and in combination with fluconazole was investigated by the classical broth microdilution method and the time-killing curve method. The QRT-PCR method was used to determine gene expressions changes of mitochondrial respiratory chain enzymes, drug efflux pumps and drug target enzymes in Candida glabrata and Candida tropicalis after n-butylphthalide treatment. The MIC values of n-butylphthalide against Candida glabrata and Candida tropicalis ranged from 16 to 64 μg·mL-1. The FICI value of the combination of n-butylphthalide and fluconazole against drug-resistant Candida glabrata and Candida tropicalis ranged from 0.5001 to 0.5315 with partial synergism. Time-killing curves showed that 256 μg·mL-1 n-butylphthalide significantly inhibited the growth of drug-resistant colonies of Candida glabrata and Candida tropicalis, and 128 μg·mL-1 n-butylphthalide combined with 1 μg·mL-1 fluconazole had an additive effect. N-butylphthalide could alter the expression of mitochondrial respiratory chain enzymes COX1, COX2, COX3, and CYTB genes in Candida glabrata and Candida tropicalis (P< 0.05) and downregulate the expression of the drug efflux pump genes CDR1 and CDR2 in drug-resistant Candida glabrata to 3.36% and 3.65%, respectively (P<0.001), but did not affect the drug target enzyme ERG11 in drug-resistant Candida tropicalis. N-butylphthalide had antifungal activity against Candida glabrata and Candida tropicalis. N-butylphthalide improved the activity of fluconazole against drug-resistant Candida glabrata by affecting the expression of mitochondrial respiratory chain enzyme genes and reversing the high expression of drug efflux pump genes CDR1 and CDR2.

Genetic diversity of the common carp black strain population based on mtDNA (D-loop and cytb)

The common strain black carp (Cyprinus carpio var. baisenensis) is a culturally important carp strain that is raised and cultured in Guangxi Province, China. Its color reflects the interactions between the Burau people and their surrounding environment. The population of the common carp black strain was isolated and cultured in a rice-fish integration system. To explore the genetic diversity and protection of germplasm resources, we analyzed mitochondrial DNA (mtDNA) sequences, specifically the displacement loop (D-loop) and cytochrome b (Cytb), using single-nucleotide polymorphisms (SNP). We compared these sequences with those from four other local common carp populations. The study included a total of 136 adult common carps from five strain populations: the common black carp strain (HJ = 31), Jian (F = 30), Heilongjiang (H = 10), Songpu (S = 31), and Saijiang (SJ = 34). The results of the Cytb and D-loop analyses showed that the Heilongjiang carp (H) and Saijiang (SJ) populations had the highest levels of haplotype diversity (0.867 ± 0.034785) and nucleotide diversity (π = 0.0063 ± 0.000137 and 0.0093 ± 0.000411), respectively. On the other hand, the Common carp black strain population (HJ) exhibited the lowest haplotype diversity in both Cytb and D-loop, with haplotype 2 being the most commonly observed among the populations. Private haplotypes dominated the five common carp populations, which were significantly different at P<0.001. Furthermore, analyzing the coefficient of genetic differentiation (Fst), the highest genetic difference was observed between Saijiang (SJ) and Heilongjiang (H) (Fst = 0.963), whereas the lowest was observed between Songpu (S) and the Common carp black strain population (HJ) (Fst = 0.019) for the Cytb gene sequences. For the D-loop, the Common carp black strain population (HJ) and Songpu (S) (Fst = 0.7) had the highest values, and Heilongjiang (H) and Common black carp strain (HJ) had an Fst of 0.125. Additionally, the AMOVA analysis revealed a higher level of variance for the Cytb and D-loop genes, indicating lower genetic diversity within the local carp community. On the other hand, the phylogenetic tree analysis showed that the five carp populations were closely related and formed a distinct cluster. The distinct cluster of populations suggests a common ancestor or recent gene flow, possibly due to geographic proximity or migration history, and unique genetic characteristics, possibly due to adaptations or selective pressures. The results of this study provide valuable insights into the genetic diversity of the common strain black carp, which can have implications for conservation, breeding programs, evolutionary studies, and fisheries management.

Genotyping, bionomics and host-feeding behavior of Phlebotomus spp. (Diptera: Psychodidae) in Iran

The main goal of the current study was to investigate the molecular systematics and host–vector interactions of sandfly species (Phlebotomus spp.) in Iran. Sandflies were collected using CDC light traps, sticky traps, and BG-Sentinel traps at seven trapping sites in Iran during 2019 and 2020. The source of blood meals was identified using primer pairs specific to either the 16S rRNA or the mitochondrial cytochrome b (cytb) genes. A total of 736 blood-fed female sandflies were collected. Six species of the genus Phlebotomus (Ph.) and four species of the genus Sergentomyia (Se.) were identified using molecular identification techniques. The most abundant sandfly species were Ph. papatasi (38.45 %) and Se. sintoni (36.28 %). Blood meal analysis identified 19 different host species, specifically, 15 mammalian and four bird species. The most common blood meal sources were Rattus norvegicus, followed by Homo sapiens, Bos primigenius f. taurus, Ovis orientalis f. aries, Meriones persicus, Gallus gallus f. domestica, Rhombomys opimus, and Canis lupus; other host species had incidences of less than 5 %. We observed that Ph. major complex and Se. sintoni had mostly mammalophilic blood-feeding behavior, while Ph. papatasi and Ph. sergenti had mostly anthropophilic and ornithophilic blood-feeding preferences, respectively. The current results confirmed cytb is a suitable gene for regular species level identification, and that phylogenetic analyses of cytb allowed for clear demarcation of sandfly species.

Molecular authentication and discrimination between banana shrimp and Pacific white shrimp by duplex PCR-lateral flow dipstick (dPCR-LFD) system

Banana shrimp is easily substituted by Pacific white shrimp due to its relatively similar morphology and appearance. The duplex PCR-lateral flow dipstick (dPCR-LFD) system was developed to authenticate these two species based on the mitochondrial cytochrome b (Cytb) gene. Two pairs of species-specific primers were designed for dPCR amplification. The specificity was also tested toward nine marine species. The sensitivity of the method was also evaluated at the DNA range of 0.02 %-60 % of adulterated Pacific white shrimp DNA in DNA from banana shrimp. Several types of cooked samples under various conditions were also tested. The developed dPCR-LFD system could specifically differentiate banana shrimp from Pacific white shrimp. No cross-amplification was observed in the specificity test. The sensitivity test showed the detection limit of 0.02 % (0.01 ng) of Pacific white shrimp DNA. The cooking processes including boiling, frying, and steaming did not show any effect on the efficiency of the dPCR-LFD system. This technique could be therefore implemented for the authentication of banana shrimp and Pacific white shrimp products as well as further use for other seafood frauds.

Features of the evolutionary transition/transversion bias of birds and mammals by the CYTB gene

Differences in the nature of the transition/transversion bias of birds and mammals, as well as families with small and large body-sized species, are proven by the example of the CYTB gene. It was that for birds compared to mammals, as well as in families of small birds and mammals compared to larger ones, the frequency of transversions is significantly higher and the frequency of transitions is lower. This leads to a decrease in the transition/transversion bias and a decrease in the rate of its evolutionary compensation. The possible cause of this phenomenon is the greater intensity of individual metabolism and the resulting increase in mutation rates in birds and small species. Exceptions are extremely small species that are characterized by a state of hypothermia. The high level of metabolism and mutability explains the richness of bird species, as well as the highest activity of speciation in small organisms. In addition, the transition/trans- version bias should be considered as a reliable integral indicator of individual metabolic intensity at the family level.

Enterocytozoon bieneusi in Wild Rats and Shrews from Zhejiang Province, China: Occurrence, Genetic Characterization, and Potential for Zoonotic Transmission.

Globally, Enterocytozoon bieneusi has been detected in humans and various animal hosts. Wild rats and shrews have the potential to act as carriers of E. bieneusi, facilitating the parasite's transmission to humans and domestic animals. We aimed to investigate the prevalence of E. bieneusi in 652 wild rats and shrews from Zhejiang Province, China, by amplifying the internal transcribed spacer (ITS) region of rDNA through polymerase chain reaction (PCR). To determine animal species, we amplified the Cytochrome b (Cyt-b) gene in their fecal DNA using PCR. Furthermore, we determined the genotype of E. bieneusi by amplifying the ITS region of rDNA through PCR. Genetic traits and zoonotic potential were evaluated using similarity and phylogenetic analyses. Suncus murinus (n = 282) and five rat species, Rattus losea (n = 18), Apodemus agrarius (n = 36), Rattus tanezumi (n = 86), Rattus norvegicus (n = 155), and Niviventer niviventer (n = 75), were identified. The average infection rate of E. bieneusi was 14.1% (92/652) with 18.1% (51/282) in S. murinus and 11.1% (41/370) in rats (27.8% in R. losea, 22.2% in A. agrarius, 10.5% in R. tanezumi, 8.4% in R. norvegicus, and 8.0% in N. niviventer). Thirty-three genotypes were identified, including 16 known genotypes. The most commonly known genotypes were HNR-VI (n = 47) and Peru11 (n = 6). Type IV, KIN-1, SHW7, and HNPL-II were each found in two samples, while Macaque4, CH5, K, Henan-III, Henan-V, HNP-II, HNPL-I, HNPL-III, HNHZ-II, and HNHZ-III were each found in one sample. Additionally, 17 novel genotypes were discovered: WZR-VIII (n = 5), WZR-I to WZR-VII, WZR-IX to WZR-XII, and WZSH-I to WZSH-V (n = 1 each). Those 33 genotypes were divided into three groups: Group 1 (n = 25), Group 2 (n = 3), and Group 13 (n = 5). The initial report underscores the extensive occurrence and notable genetic diversity of E. bieneusi in wild rats and shrews from Zhejiang province, China. These results suggest that these animals play a pivotal role in the transmission of E. bieneusi. Furthermore, animals carrying the zoonotic genotypes of E. bieneusi pose a serious threat to residents.

Differential Mitochondrial Genome Expression of Three Sympatric Lizards in Response to Low-Temperature Stress.

Ecological factors related to climate extremes have a significant influence on the adaptability of organisms, especially for ectotherms such as reptiles that are sensitive to temperature change. Climate extremes can seriously affect the survival and internal physiology of lizards, sometimes even resulting in the loss of local populations or even complete extinction. Indeed, studies have shown that the expression levels of the nuclear genes and mitochondrial genomes of reptiles change under low-temperature stress. At present, the temperature adaptability of reptiles has rarely been studied at the mitochondrial genome level. In the present study, the mitochondrial genomes of three species of lizards, Calotes versicolor, Ateuchosaurus chinensis, and Hemidactylus bowringii, which live in regions of sympatry, were sequenced. We used RT-qPCR to explore the level of mitochondrial gene expression under low-temperature stress, as compared to a control temperature. Among the 13 protein-coding genes (PCGs), the steady-state transcript levels of ND4L, ND1, ATP6, and COII were reduced to levels of 0.61 ± 0.06, 0.50 ± 0.08, 0.44 ± 0.16, and 0.41 ± 0.09 in C. versicolor, respectively, compared with controls. The transcript levels of the ND3 and ND6 genes fell to levels of just 0.72 ± 0.05 and 0.67 ± 0.05 in H. bowringii, compared with controls. However, the transcript levels of ND3, ND5, ND6, ATP6, ATP8, Cytb, and COIII genes increased to 1.97 ± 0.15, 2.94 ± 0.43, 1.66 ± 0.07, 1.59 ± 0.17, 1.46 ± 0.04, 1.70 ± 0.16, and 1.83 ± 0.07 in A. chinensis. Therefore, the differences in mitochondrial gene expression may be internally related to the adaptative strategy of the three species under low-temperature stress, indicating that low-temperature environments have a greater impact on A. chinensis, with a small distribution area. In extreme environments, the regulatory trend of mitochondrial gene expression in reptiles is associated with their ability to adapt to extreme climates, which means differential mitochondrial genome expression can be used to explore the response of different lizards in the same region to low temperatures. Our experiment aims to provide one new research method to evaluate the potential extinction of reptile species in warm winter climates.

Molecular detection of Lotmaria passim in intestine of Japanese honeybees (Apis cerana japonica)

ABSTRACTCrithidia mellificae and Lotmaria passim are trypanosomatids that infect honeybees, and many studies suggest that both parasites contribute to the decline in the honeybee population. The Japanese honeybee (Apis cerana japonica) is a native honeybee that inhabits various areas of Japan and is one of the most important pollinators. Both parasites have been well studied in the western honeybee (Apis mellifera) but have been poorly investigated in A. c. japonica. The present study investigated the presence of C. mellificae and L. passim in one feral and six managed A. c. japonica colonies at three different institutes. Five out of seven colonies were polymerase chain reaction‐positive for L. passim (71.4%); however, C. mellificae was not detected. Four of the five colonies were positive in both the midgut and hindgut, but one was positive only in the hindgut. A homology search and phylogenetic tree of the glycosomal glyceraldehyde‐3‐phosphate dehydrogenase (gGAPDH) and cytochrome b (Cytb) genes obtained in this study indicated that the sequences belonged to L. passim. The results of this study emphasize concerns regarding the health of this ecologically important pollinator and the importance of further investigation of the prevalence of L. passim in A. c. japonica in a wider area of Japan.

Aptly chosen, effectively emphasizing the action and mechanism of antimycin A1.

Rhizoctonia solani Kühn, a plant pathogenic fungus that can cause diseases in multiple plant species is considered one of the common and destructive pathogens in many crops. This study investigated the action of antimycin A1, which was isolated from Streptomyces AHF-20 found in the rhizosphere soil of an ancient banyan tree, on Rhizoctonia solani and its mechanism. The inhibitory effect of antimycin A1 on R. solani was assessed using the comparative growth rate method. The results revealed that antimycin A1 exhibited a 92.55% inhibition rate against R. solani at a concentration of 26.66 μg/mL, with an EC50 value of 1.25 μg/mL. To observe the impact of antimycin A1 on mycelial morphology and ultrastructure, the fungal mycelium was treated with 6.66 μg/mL antimycin A1, and scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were employed. SEM analysis demonstrated that antimycin A1 caused mycelial morphology to become stripped, rough, and folded. The mycelium experienced severe distortion and breakage, with incomplete or locally enlarged ends, shortened branches, and reduced numbers. TEM observation revealed thickened cell walls, indistinct organelle boundaries, swollen mitochondria, exosmotic substances in vesicles, slow vesicle fusion, and cavitation. Real-time quantitative PCR and enzyme activity assays were conducted to further investigate the impact of antimycin A1 on mitochondria. The physiological and biochemical results indicated that antimycin A1 inhibited complexes III and IV of the mitochondrial electron transport chain. RT-PCR analysis demonstrated that antimycin A1 controlled the synthesis of relevant enzymes by suppressing the transcription levels of ATP6, ATP8, COX3, QCR6, CytB, ND1, and ND3 genes in mitochondria. Additionally, a metabolomic analysis revealed that antimycin A1 significantly impacted 12 metabolic pathways. These pathways likely experienced alterations in their metabolite profiles due to the inhibitory effects of antimycin A1. Consequently, the findings of this research contribute to the potential development of novel fungicides.

Evidence for Selection on Mitochondrial OXPHOS Genes in the Mediterranean Killifish Aphanius fasciatus Valenciennes, 1821.

Mitochondrial oxidative phosphorylation (OXPHOS) genes are a system subject to selection under determined environmental constraints despite a neutral evolution model that has long been hypothesized for the mitochondrial genome. In this study, the sequences of ND1, Cytb, and COI OXPHOS genes were analyzed in six populations of the eurythermal and euryhaline killifish A. fasciatus, to detect non-synonymous mutations leading to amino acid changes and to check whether selection acted on them using tests of recombination and selection. The results indicate a high COI and Cytb gene diversity and a high percentage of private haplotypes in all populations. In the Greek population, non-synonymous nucleotide substitutions were observed in the N-terminal region of COI and Cytb. Positively selected sites were also found. The information we obtained from the mitochondrial DNA sequences of A. fasciatus adds to the growing data on selective pressure acting on mitochondrial DNA in non-model species. These results should be explored from the perspective of the local adaptation of eurythermal and euryhaline species and supported using experimental evidence to better understand the interplay between historical climatic events and local adaptation and how each of them contributes to shaping the genetic structure of this species.

Identification of Cervi Cornu (Cervus elaphus) and its formula granules based on PCR-RFLP

Cervi Cornu is the ossified antler, or the base antler that falls off in the spring of the following year after the pilose antler is sawn off from Cervus elaphus or C. nippon, as a precious traditional Chinese medicine, has been recognized for its medicinal value and widely used in clinical practice. However, the origins of Cervi Cornu are miscellaneous, and Cervi Cornu is even mixed with adulterants in the market. Currently, there is a shortage of ways to identify Cervi Cornu and no standard to control the quality of Cervi Cornu. So it is valuable to develop a way to effectively identify Cervi Cornu from the adulterants. In this study, the differences in the mitochondrial barcode cytochrome b(Cytb) gene sequences of C. elaphus, C. nippon and their related species were compared and the specific single nucleotide polymorphism(SNP) sites on the Cytb sequences of Cervi Cornu were screened out. According to the screened SNPs, Cervi Cornu-specific primers dishmy-F and dishmy-R were designed. The PCR system was established and optimized, and the tolerance and feasibility of Taq polymerases and PCR systems affecting the repeatability of the PCR method were investigated. The amplification products of C. elaphus and C. nippon were digested using the restriction enzyme MseⅠ. The results showed that after electrophoresis of the product from PCR with the annealing temperature of 56 ℃ and 35 cycles, a single specific band at about 100 bp was observed for C. elaphus samples, and the product of C. elaphus samples was 60 bp shorter than that of C. nippon samples. There was no band for adulterants from other similar species such as Alces alces, Rangifer tarandus, Odocoileus virginianus, O. hemionus, Cap-reolus pygargus, Przewalskium albirostis and negative controls. The polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) method established in this study can quickly and accurately identify Cervi Cornu originated from C. elaphus in crude drugs, standard decoctions, and formula granules, and distinguish the origins of Cervi Cornu products, i.e., C. nippon and similar species. This study can be a reference for other studies on the quality standard of other formula granules of traditional Chinese medicines.

Differences in haemosporidian parasite prevalence and diversity in migratory and resident birds of prey species revealed by a non-invasive sampling method.

Avian haemosporidian parasites are widespread globally and infect numerous wild bird species. However, they have primarily been studied in passerine birds. Accordingly, the prevalence and diversity of these parasites in birds of prey remain understudied. This lack of research is primarily due to the international protection status of many of these species, their sparse distribution across ecosystems and difficulty to capture in the wild. The aim of this study was to examine the prevalence and diversity of haemosporidian parasites in two species of birds of prey, namely white-tailed eagle and lesser spotted eagle. To achieve this, a non-invasive approach was employed, involving the extraction of DNA from blood spots present in moulted feathers. Freshly moulted feathers were collected from the ground under nests or within the nests of these birds during the breeding season. A visible blood spot located at the junction of the calamus and rachis was removed and fixed in SET buffer for molecular analysis. The identification of haemosporidian parasites (Plasmodium, Haemoproteus and Leucocytozoon) was conducted using PCR-based methods. Overall, parasite DNA was successfully detected in shed feathers that were kept in their original form at least decade prior to analysis. Among the studied individuals, resident white-tailed eagles showed significantly lower infection rates of haemosporidian parasites compared to migratory lesser spotted eagles. A total of nine genetic lineages of haemosporidian parasites were detected, with seven of them being new to science based on partial sequences of the cytb gene. Additionally, the phylogenetic relationships among these new lineages and previously described ones were established. These findings highlight the suitability of non-invasive sampling for investigating the prevalence and diversity of haemosporidian parasites in wild birds of prey populations. Moreover, this approach holds promise for studying other challenging-to-reach and protected bird species. According to our research, there is a greater chance of finding haemosporidian parasites in freshly gathered feathers.

EDNA Metabarcoding: Gaps of Publicly Available Reference Databases of Freshwater Vertebrates in Sri Lanka

Environmental DNA (eDNA) has become a rapidly emerging technology during the last decade. Although eDNA applications are used worldwide, none of the eDNA metabarcoding studies have been conducted solely in Sri Lanka yet. Detection rate of species through DNA metabarcoding technique varies with the availability of gene sequences in reference databases. Therefore, the main objective of this study was to assess the data gaps of local species in two selected reference databases (the NCBI database and the BOLD database). Local freshwater vertebrates were considered for this analysis by dividing them into three major categories as freshwater fish (97 species), freshwater amphibians (40 species) and freshwater reptiles (7 species). The availability of COI gene sequence and CytB gene sequence of each species were recorded. In the NCBI database, reference gene sequences were available only for 17.53% (17 species) of freshwater fish, 7.50% (3 species) of freshwater amphibians and 14.29% (1 species) of freshwater reptiles. In the BOLD database, reference gene sequences were available for 52.58% (51 species) of freshwater fish, 17.50% (7 species) of freshwater amphibians and 42.86% (3 species) of freshwater reptiles. Endemic species were analyzed separately. This revealed the availability of reference gene sequences for 4.92% (3 species) of endemic freshwater fish in the NCBI database. However, none of the reference gene sequence was found for endemic freshwater amphibians and reptiles. In the BOLD database, reference gene sequences were available for 36.07% (22 species) of endemic freshwater fish and 33.33% (1 species) of endemic freshwater reptiles but none for endemic freshwater amphibians. According to the results, large gaps of publicly available reference CO1 gene sequences were observed in both NCBI and BOLD databases. Not only the availability of reference gene sequences, but also the submitted countries of those sequences were also analyzed as a part of this study. Asia has contributed the most according to the country-vice analysis. Among them, many are in the South Asian region. Therefore, this study highlights the need of expanding reference databases for greater local representation as well as the importance of forging collaborations with the developed countries in the same region in order to increase international cooperation and funding to carry on further studies on neglected taxa and under‐represented taxa in Sri Lanka. &#x0D; Keywords: eDNA (environmental DNA), Metabarcoding, Reference databases, Reference gene sequences

Nanoplate-based digital PCR for highly sensitive pork DNA detection targeting multi-copy nuclear and mitochondrial genes

The inclusion of ingredients derived from pigs in highly processed consumer products poses a significant challenge for DNA-targeted analytical enforcement, which could be overcome by using digital PCR. However, most species detection methods use digital PCR to target single-copy nuclear genes, which limits their sensitivity. In this work, we examined the performance of a nanoplate-based digital PCR method that targets multi-copy nuclear (MPRE42) and mitochondrial (Cytb) genes. Poor separation of positive and negative partitions, as well as a ‘rain effect’ were obtained in the porcine-specific MPRE42 assay. Among the optimization strategies examined, the inclusion of restriction enzymes slightly improved the separation of positive and negative partitions, but a more extensive ‘rain effect’ was observed. The high copy number of the MPRE42 amplicon is hypothesized to contribute to the saturation of the positive signal. In contrast, the porcine-specific Cytb assay achieved perfect separation of positive and negative partitions with no ‘rain effect’. This assay can detect as little as 0.4 pg of pork DNA, with a sensitivity of 0.05% (w/w) in a pork-chicken mixture, proving its applicability for detecting pork in meat and meat-based products. For the MPRE42 assay, potential applications in highly degraded products such as gelatin and lard are anticipated.

Detection and Genotyping of Trypanosoma cruzi Samples in Species of Genus Rhodnius from Different Environments in the Brazilian Amazon.

Background: In the Amazon region, several species of triatomines occur in the natural environments. Among them, species of the genus Rhodnius are a risk to human populations due to their high rates of infection with Trypanosoma cruzi. The aim of this study was to identify the T. cruzi genotypes in Rhodnius specimens and their relationship with sylvatic hosts from different environments in the Brazilian Amazon. Methods: A total of 492 triatomines were collected from the municipalities of Monte Negro, Rondônia state, and Humaitá, Amazonas state, 382 of them being nymphs and 110 adults. Genotyping of T. cruzi in six discrete typing units (DTUs) was performed using conventional multilocus PCR. The triatomines that were positive for T. cruzi and engorged with blood were also targeted for amplification of the cytochrome B (cytB) gene to identify bloodmeal sources. Results: Of the 162 positive samples, the identified DTUs were TcI (87.65%) and TcIV (12.35%). It was observed that 102 specimens were engorged with a variety of bloodmeals. Triatomines infected with TcI were associated with DNA of all identified vertebrates, except Plecturocebus brunneus. TcIV was detected in triatomines that fed on Coendou prehensilis, Didelphis marsupialis, Mabuya nigropunctata, P. brunneus, Pithecia irrorata, Sapajus apella, and Tamandua tetradactyla. Conclusion: Results highlight the need to understand the patterns of T. cruzi genotypes in Rhodnius spp. and their association with sylvatic hosts to better elucidate their role in the transmission of Chagas disease in the Amazon region.

Diversity Analysis of Asian Clam Collected from Selected Rivers in Kelantan

The phylogenetic study was conducted to reveal the types of Corbicula species located on a selected river in Kelantan. The morphotypes and morphometric features were used to analyse the phenotypic analysis, and genotypes were examined exploiting the mitochondrial DNA cytochrome b (cytb) gene. A unique morphotype round (R) was discovered for every examined sample. Furthermore, there were significant differences (p&lt;0.05) in the morphometric features (umbo length, UL; shell width, SW; and shell height, SH) between the C. fluminea populations. The mtDNA cytb sequences are used in the genotyping analysis to confirm that the assessed Corbicula is C. fluminea. Three genotypes were identified in the mtDNA cytb phylogenetic tree. In geographically isolated populations, phylogenetic trees showed polymorphism and minimal genetic variability, which aligned with the phenotypic characterisation. The results clarify the differences among C. fluminea and imply that Corbicula sp. in Kelantan originated from a single species. These findings are significant not only for biologists or those involved in ecology and ecosystems but also for the fisheries sector as it serves as the basis for breeding purposes, overcome the extinction, and enhancing the economy of the inland fishermen in Kelantan.

Molecular characterization of Cryptosporidium in wild rodents from the Inner Mongolian Autonomous Region and Liaoning Province, China: assessing host specificity and the potential for zoonotic transmission.

Wild rodents are key hosts for Cryptosporidium transmission, yet there is a dearth of information regarding their infection status in the Inner Mongolian Autonomous Region and Liaoning Province of China. Therefore, the present study was conducted to determine the prevalence and genetic characteristics of Cryptosporidium among wild rodents residing in these two provinces. A total of 486 rodents were captured, and fresh feces were collected from each rodent's intestine for DNA extraction. Species identification of rodents was performed through PCR amplification of the vertebrate cytochrome b (cytb) gene. To detect the presence of Cryptosporidium in all fecal samples, PCR analysis and sequencing of the partial small subunit of the ribosomal RNA (rRNA) gene were performed. Four species of rodents were identified: Rattus norvegicus, Mus musculus, Apodemus agrarius, and Cricetulus barabensis. Positive results for Cryptosporidium were obtained for 9.2% (18/195), 6.6% (7/106), 5.6% (5/89), and 6.3% (6/96) of these rodents, respectively, with an average infection rate of 7.4% (36/486). The identification revealed the presence of five Cryptosporidium species, C. ubiquitum (n = 8), C. occultus (n = 5), C. muris (n = 2), C. viatorum (n = 1), and C. ratti (n = 1), along with two Cryptosporidium genotypes: Rat genotype III (n = 10) and Rat genotype IV (n = 9). Based on the molecular evidence presented, the wild rodents investigated were concurrently infected with zoonotic (C. muris, C. occultus, C. ubiquitum and C. viatorum) as well as rodent-adapted (C. ratti and Rat genotype III and IV) species/genotypes, actively participating in the transmission of cryptosporidiosis.

Fungicide-Resistant Botrytis in Forest Nurseries May Impact Disease Control in Norway Spruce.

Gray mold, caused by Botrytis spp., is a serious problem in Norway spruce seedling production in forest nurseries. From 2013 to 2019, 125 isolates of Botrytis were obtained from eight forest nurseries in Norway: 53 from Norway spruce seedlings, 16 from indoor air, 52 from indoor surfaces, and four from weeds growing close to seedlings. The majority of isolates were identified as B. cinerea, and over 60% of these were characterized as Botrytis group S. B. pseudocinerea isolates were obtained along with isolates with DNA sequence similarities to B. prunorum. Fungicide resistance was assessed with a mycelial growth assay, and resistance was found for the following: boscalid (8.8%), fenhexamid (33.6%), fludioxonil (17.6%), pyraclostrobin (36.0%), pyrimethanil (13.6%), and thiophanate-methyl (50.4%). Many isolates (38.4%) were resistant to two to six different fungicides. A selection of isolates was analyzed for the presence of known resistance-conferring mutations in the cytb, erg27, mrr1, sdhB, and tubA genes, and mutations leading to G143A, F412S, ΔL497, H272R, and E198A/F200Y were detected, respectively. Detection of fungicide resistance in Botrytis from Norway spruce and forest nursery facilities reinforces the necessity of employing resistance management strategies to improve control and delay development of fungicide resistance in the gray mold pathogens.[Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Molecular investigation of Blastocystis sp. infections in wild rodents from the Inner Mongolian Autonomous Region and Liaoning province, China: High prevalence and dominance of ST4.

Wild rodents are key carriers of various human pathogens, including Blastocystis spp. Our study aimed to assess the prevalence and genetic characteristics of Blastocystis among wild rodents in the Inner Mongolian Autonomous Region and Liaoning Province of China. From November 2023 to February 2024, 486 rodents were captured in these regions. Fresh feces were collected from the intestines of each rodent for the isolation of DNA and PCR amplification of the vertebrate cytochrome b (cytb) gene to identify rodent species. Subsequently, PCR analysis and sequencing of the partial small subunit of the ribosomal RNA (rRNA) gene were utilized to detect Blastocystis in all fecal samples. Of the total samples, 27.4% (133/486) were found to be Blastocystis positive. The results revealed the presence of four species of rodents infected with Blastocystis, 32.3% (63/195) in Rattus norvegicus, 15.1% (16/106) in Mus musculus, 20.2% (18/89) in Apodemus agrarius, and 37.5% (36/96) in Cricetulus barabensis. Sequence analysis confirmed the existence of five Blastocystis subtypes: ST1 (n=4), ST2 (n=2), the ST4 (n=125, the dominant subtype), ST10 (n=1), and a novel ST (n=1). The identified zoonotic subtypes (ST1, ST2, ST4, and ST10) highlight the possible role played by wild rodents in the transmission of Blastocystis to humans, thereby elevating the chances of human infection. Meanwhile, the discovery of novel sequences also provides new insights into the genetic diversity of this parasite.

Monilia and Monilinia : In silico genetic analysis of plant pathogenic fungi

Species of phytopathogenic fungi Monilia [J. Hill, 1751] and Monilinia [E.E. Honey, 1936] of the phylum Ascomycota have two host plant families of stone- and pome fruit trees of Rosaceae; and bushes of Ericaceae (e.g., blueberry, cranberry, heather, etc.). They infect both plants, and human (i.e., moniliasis). Due to the lack of sexual developmental stage (i.e., imperfect fungi; anamorphic fungi) the species of the order Moniliales spread by vegetative spores (i.e., conidia) and not by sexual (i.e., meiotic) ascospores. Here we report in silico data mining carried out by analyses of DNA, RNA, and protein sequences to reveal genetic distances in -and-among Monilia and Monilinia species. Sequences of the (+)ssRNA genomes of mitoviruses, which hyper parasitize the fungi cells, which parasitize the plant cells, during the ecocycles, are also analyzed. Genome size analysis of Monilinia were found to be between 30-55 x 106 bp; the cytochrome-b (cytb) genes (1543 bp of mtDNA); the transcribed cytb-mRNAs; the translated cytb proteins (381-391 aa); and the (+)ssRNA genomes of mitoviruses have showed that Botrytis cinerea (‘grey mold’) has the closest molecular similarity to Monilia and Monilinia clade (‘brown rot’). The cladogram of transcribed cytb-mRNAs (1175 nt) grouped Monilinia vaccinii-corymbosi in a distinct clade, which indicate a far genetic linkage. The application of mitovirus-infected hypovirulent phytopathogenic fungi for plant protection (i.e., virocontrol / biocontrol) is discussed.