Molecular prevalence of Tetratrichomonas gallinarum and Trichomonas gallinae in three domestic free-range poultry breeds in Anhui Province, China.

Tetratrichomonas gallinarum and Trichomonas gallinae can colonize the alimentary tract of domestic birds. However, little information is available on the epidemiology of the two trichomonad species in domestic free-range poultry in China. In this study, the occurrence and genetic characteristic of T. gallinarum and T. gallinae among free-range chickens, ducks, and geese in Anhui Province, China, were investigated. The 1910 fecal samples collected from 18 free-range poultry farms throughout Anhui Province were examined for the presence of T. gallinarum and T. gallinae by PCR and sequence analysis of the small subunit (SSU) rRNA gene of T. gallinarum and ITS1-5.8S-ITS2 sequence of T. gallinae. The overall occurrence of T. gallinarum in poultry was 1.2% (22/1910), with infection rates of 2.1% (17/829) in chickens, 0.2% (1/487) in ducks, and 0.7% (4/594) in geese. The constructed phylogeny tree using the concatenated ITS1-5.8S-ITS2 region and SSU rRNA indicated the T. gallinarum isolates detected in this study were closely related to previously defined genogroups A, D, and E, respectively. Nine (0.5%) fecal samples were positive for T. gallinae, with infection rates of 0.8% (7/829) in chickens, 0.4% (2/487) in ducks, and 0% (0/594) in geese. Sequence and phylogenetic analysis showed that four T. gallinae ITS1-5.8S-ITS2 sequences obtained from chicken feces and one duck fecal sample belonged to genotype ITS-OBT-Tg-1. This is the first report of the prevalence and genetic characterization of T. gallinarum and T. gallinae in free-range chickens, ducks, and geese in China.

Transmission of the wMel Wolbachia strain is modulated by its titre and by immune genes in Drosophila melanogaster (Wolbachia density and transmission).

Wolbachia are common intracellular endosymbionts of arthropods, but the interactions between Wolbachia and arthropods are only partially understood. The fruit fly Drosophila melanogaster is a model insect for understanding Wolbachia-host interactions. Here the native wMel strain of D. melanogaster was isolated and then different initial titres of wMel were artificially transferred back into antibiotics-treated fruit flies. Our purpose was to examine the interactions between the injected wMel in a density gradient and the recipient host during trans-generational transmission. The results showed that the trans-generational transmission rates of wMel and titres of wMel exhibited a fluctuating trend over nine generations, and the titres of wMel displayed a similar fluctuating trans-generational trend. There was a significant positive correlation between the transmission rate and the titre of wMel. Reciprocal crossings between wMel-transinfected and uninfected fruit flies revealed that wMel could induce cytoplasmic incompatibility (CI) at different initial titres, but the intensity of CI was not significantly correlated with the initial titre of wMel. Quantitative PCR analysis showed that the immune genes Drsl5 and Spn38F displayed a significant transcriptional response to wMel transfection, with an obvious negative correlation with the titre of wMel at the 3rd and 4th generations. Furthermore, RNA interference-mediated knockdown of Drsl5 and Spn38F elicited a drastic increase in the titre of wMel. In combination, our study suggests that the trans-generational transmission of wMel is modulated by its density, and the immune genes are involved in the regulation of Wolbachia density.

Molecular prevalence and characterization of Cryptosporidium in domestic free-range poultry in Anhui Province, China.

Free-range chickens might mediate the spread of Cryptosporidium oocysts to humans and other animals. Few studies have evaluated the prevalence of Cryptosporidium species in domestic free-range poultry in China. Here, we characterized the prevalence and distribution of species and genotypes of Cryptosporidium in domestic free-range chickens, ducks, and geese in Anhui Province, China. A total of 1910 fresh fecal samples from three poultry species were examined from 18 free-range poultry farms by nested PCR and analysis of the Cryptosporidium SSU rRNA gene. The overall prevalence of Cryptosporidium species was 2.9% (55/1910), with infection rates of 1.3% (11/829) in chickens, 7.3% (36/487) in ducks, and 1.4% (8/594) in geese. C. baileyi (0.6%), C. meleagridis (0.2%), C. galli (0.2%), and C. xiaoi-like genotype (0.2%) were identified in chickens, and only C. baileyi was identified in ducks and geese, with infection rates of 7.4% and 1.3%, respectively. C. baileyi was the most prevalent species. Sequencing of the GP60 gene revealed that the C. meleagridis isolates belonged to the IIIbA26G1R1b subtype. This is the first study to document C. galli and C. xiaoi-like genotype in domestic free-range chickens in China. These findings expand the range of avian hosts known for Cryptosporidium and highlight the need for additional studies to characterize the diversity of Cryptosporidium in avian species.

The Intruding Wolbachia Strain from the Moth Fails to Establish Itself in the Fruit Fly Due to Immune and Exclusion Reactions.

Wolbachia is capable of regulating host reproduction, and thus of great significance in preventing the spread of insect-borne diseases and controlling pest insects. The fruit fly Drosophila melanogaster is an excellent model insect for understanding Wolbachia-host interactions. Here we artificially transferred the wCcep strain from the rice moth Corcyra cephalonica into D. melanogaster by microinjection. Crossing experiments indicated that wCcep could induce a high level of CI in the phylogenetically distant host D. melanogaster and imposed no negative fitness costs on host development and fecundity. Based on quantitative analysis, the titres of wCcep and the native wMel strain were negatively correlated, and wCcep could only be transmitted in the novel host for several generations (G0 to G4) after transinfection. Transcriptome sequencing indicated that the invading wCcep strain induced a significant immune- and stress-related response from the host. An association analysis between the expression of immune genes attacin-D/edin and the titre of Wolbachia by linear regression displayed a negative correlation between them. Our study suggest that the intrusion of wCcep elicited a robust immune response from the host and incurred a competitive exclusion from the native Wolbachia strain, which resulted in the failure of its establishment in D. melanogaster.

Prevalence of Tetratrichomonas buttreyi and Pentatrichomonas hominis in yellow cattle, dairy cattle, and water buffalo in China.

The trichomonad species Tetratrichomonas buttreyi and Pentatrichomonas hominis have been reported in the bovine digestive tract in only a few studies, and the prevalence and pathogenicity of these two protists in cattle herds remain unknown. In this study, the prevalence of T. buttreyi and P. hominis in yellow cattle, dairy cattle, and water buffalo in Anhui Province, China, was determined with a PCR analysis of the small subunit ribosomal RNA genes. The overall infection rates for T. buttreyi and P. hominis were 8.1% and 5.4%, respectively. Double infections were found in 15 (1.6%) samples from four farms. The prevalence of P. hominis in cattle with abnormal feces was significantly higher than that in cattle with normal feces (χ2 = 13.0, p < 0.01), and the prevalence of T. buttreyi in the northern region of Anhui Province was also significantly higher than that in the mid region (χ2 = 16.6, p < 0.01). Minor allelic variations were detected in the T. buttreyi isolates from cattle in this study, as in other hosts in previous studies. Morphological observations, together with the PCR analysis, demonstrated that the trichomonads isolated in this study were P. hominis. The presence of T. buttreyi and P. hominis indicated that cattle are natural hosts of these two trichomonads and could be a potential source of P. hominis infections in humans and other animal hosts.

The first report of Cryptosporidium testudinis in Chinese alligators (Alligator sinensis) in China.

Several Cryptosporidium species that infect reptiles, especially squamates, are well described, but there is limited data about Cryptosporidium species infecting crocodilians. In this study, we assess the occurrence of intestinal parasites using traditional microscopic examination and describe the prevalence and Cryptosporidium species in the captive-bred Chinese alligators (Alligator sinensis) in eastern China using molecular methods. The results of microscopic examination showed that no intestinal parasites were detected among the 491 fecal samples examined from the Chinese alligators. The overall prevalence for Cryptosporidium was 0.41% (2/491) by PCR detection using the SSU rRNA locus. Sequence and phylogenetic analysis of the SSU rRNA, COWP, and actin genes revealed the presence of Cryptosporidium testudinis, which has been isolated primarily from chelonians. This is the first detection of the specific DNA of C. testudinis in the feces of the Chinese alligator. This study expands our knowledge of the Cryptosporidium species involved in crocodiles, and more extensive studies are necessary to confirm the validity of C. testudinis in crocodiles.

Diagnostic Utility of Interferon-Gamma Release Assay in Tuberculous Lymphadenitis.

Objective The aim of this study was to evaluate the diagnostic performance of T-SPOT.TB for tuberculous lymphadenitis. Methods Suspected tuberculous lymphadenitis patients between September 2010 and September 2018 who had both peripheral blood T-SPOT.TB test and lymph node biopsy were retrospectively enrolled in this study. The cutoff value of T-SPOT.TB test for peripheral blood was set as 24 spot forming cell (SFC)/10 6 periphreral blood monocyte cell (PBMC) according to the instruction of testing kits. The gold standard for diagnosis of TBL was the combination of microbiology results, histopathology results and patient's response to anti-TB treatment. Diagnostic efficacy of T-SPOT.TB was evaluated, including sensitivity, specificity, accuracy, predictive values, and likelihood ratio. Results Among 91 patients who met the inclusion criteria, we excluded 8 cases with incomplete clinical information and 6 cases who lost to follow-up. According to the gold standard, there were 37 cases of true TBL (9 confirmed TBL and 28 probable TBL), 30 cases of non-TBL, and 10 cases of clinically indeterminate diagnosis who were excluded from the final analyses. The T-SPOT.TB tests yielded 43 cases of positive response and 24 cases of negative response. The sensitivity, specificity, accuracy, positive predictive value (PPV), negative predictive value (NPV), positive likelihood ratio (PLR) and negative likelihood ratio (NLR) of peripheral blood T-SPOT.TB for diagnosing TBL were 89.2%, 66.7%, 79.1%, 76.7%, 83.3%, 2.68 and 0.16, respectively. The number of SFCs of T-SPOT.TB in TBL patients [432(134-1264)/10 6 PBMCs] was higher than that in non-TBL patients [0 (0-30) /10 6PBMCs] with a significant difference (Z=-5.306, P <0.001).Conclusion T-SPOT.TB is a rapid and simple diagnostic test for TBL with a high sensitivity and negative predictive value.

Bioactivities of a New Pyrrolidine Alkaloid from the Root Barks of Orixa japonica.

A new pyrrolidine alkaloid named (Z)-3-(4-hydroxybenzylidene)-4-(4-hydroxyphenyl)-1-methylpyrrolidin-2-one was isolated from the ethanol extract of the root barks of Orixa japonica. The structure of the new alkaloid was elucidated on the basis of NMR and MS analysis. The compound exhibited larvicidal activity against the fourth instar larvae of Aedes aegypti (LC50 = 232.09 µg/mL), Anopheles sinensis (LC50 = 49.91 µg/mL), and Culex pipiens pallens (LC50 = 161.10 µg/mL). The new alkaloid also possessed nematicidal activity against Bursaphelenchus xylophilus (LC50 = 391.50 µg/mL) and Meloidogynein congnita (LC50 = 134.51 µg/mL). The results indicate that the crude ethanol extract of O. japonica root barks and its isolated pyrrolidine alkaloid have potential for development into natural larvicides and nematicides.

Laboratory Evaluation of Larvicidal Activity of the Essential oil of Allium tuberosum Roots and its Selected Major Constituent Compounds Against Aedes albopictus (Diptera: Culicidae).

The aim of this research was to evaluate larvicidal activity of the essential oil of Allium tuberosum Rottler ex Sprengle roots and its constituents against larval mosquitoes (Aedes albopictus Skuse). Essential oil of A. tuberosum was obtained by hydrodistillation and analyzed by gas chromatography and gas chromatography-mass spectrometry. The major constituents of the oil were found to be sulfur-containing compounds, including allyl methyl trisulfide (35.19%), diallyl disulfide (28.31%), diallyl trisulfide (20.91%), and dimethyl trisulfide (12.33%). The essential oil of A. tuberosum exhibited larvicidal activity against the fourth-instar larvae of Ae. albopictus, with an LC50 value of 18 µg/ml. The constituents compounds-diallyl trisulfide (LC50 = 4 µg/ml) and diallyl disulfide (LC50 = 6 µg/ml)-possessed stronger larvicidal activity than allyl methyl trisulfide (LC50 = 27 µg/ml) and dimethyl trisulfide (LC50 = 35 µg/ml) against the fourth-instar larvae of Ae. albopictus. The results indicated that the essential oil of A. tuberosum and its major constituents have good potential as a source for natural larvicides.

Evaluation of Contact Toxicity and Repellency of the Essential Oil of Pogostemon cablin Leaves and Its Constituents Against Blattella germanica (Blattodae: Blattelidae).

The aim of this research was to evaluate contact toxicity and repellency of the essential oil of Pogostemon cablin (Blanco) Bentham leaves against German cockroaches (Blattella germanica) (L.) and to isolate any active constituents. Essential oil of P. cablin leaves was obtained by hydrodistillation and analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Twenty-three components were identified in the essential oil, and the main constituents were patchoulol (41.31%), pogostone (18.06%), α-bulnesene (6.56%), caryophyllene (5.96%), and seychellene (4.32%). Bioactivity-directed chromatographic separation of the essential oil led to the isolation of pogostone, patchoulol, and caryophyllene as active compounds. The essential oil of P. cablin leaves exhibited acute toxicity against male B. germanica adults with an LC50 value of 23.45 µg per adult. The constituent compound, pogostone (LC50 = 8.51 µg per adult) showed stronger acute toxicity than patchoulol (LC50 = 207.62 µg per adult) and caryophyllene (LC50 = 339.90 µg per adult) against the male German cockroaches. The essential oil of P. cablin leaves and the three isolated constituents exhibited strong repellent activity against German cockroaches at a concentration of 5 ppm. The results indicated that the essential oil of P. cablin leaves and its major constituents have good potential as a source for natural insecticides and repellents.

Larvicidal activity of the essential oil of Youngia japonica aerial parts and its constituents against Aedes albopictus.

The aim of this study was to evaluate the larvicidal activity of the essential oil of Youngia japonica aerial parts against the larvae of Aedes albopictus and to isolate any active compounds from the oil. Gas chromatography-mass spectrometry (GC-MS) analyses revealed the presence of 31 compounds, with menthol (23.53%), α-asarone (21.54%), 1,8-cineole (5.36%), and caryophyllene (4.45%) as the major constituents. Bioactivity-directed chromatographic separation of the oil led to the isolation of menthol and α-asarone as active compounds. The essential oil of Y. japonica exhibited larvicidal activity against the fourth instar larvae of A. albopictus with an LC50 value of 32.45 µg/mL. α-Asarone and menthol possessed larvicidal activity against the fourth instar larvae of A. albopictus with LC50 values of 24.56 µg/mL and 77.97 µg/mL, respectively. The results indicate that the essential oil of Y. japonica aerial parts and the two constituents can be potential sources of natural larvicides.

Repellent and insecticidal effects of the essential oil of Kaempferia galanga rhizomes to Liposcelis bostrychophila (Psocoptera: Liposcelidae).

The aim of this research was to determine chemical composition and repellent and insecticidal activities of the essential oil of Kaempferia galanga L. rhizomes against the booklouse, Liposcelis bostrychophila Badonnel, and to isolate insecticidal or repellent constituents from the oil. The essential oil was obtained by hydrodistillation and analyzed by gas chromatography-mass spectrometry. Twenty-eight components of the oil were identified. The major compounds in the oil were ethyl-rho-methoxycinnamate (38.6%), ethyl cinnamate (23.2%), 1,8-cineole (11.5%), trans-cinnamaldehyde (5.3%), and borneol (5.2%). Based on bioactivity-guided fractionation, four active constituents were isolated from the oil and identified as 1,8-cineole, ethyl cinnamate, ethyl rho-methoxycinnamate, and trans-cinnamaldehyde. The essential oil exhibited contact toxicity against the booklouse with an LC50 value of 68.6 microg/cm2. Ethyl cinnamate (LC50 = 21.4 microg/cm2) exhibited stronger contact toxicity than ethyl rho-methoxycinnamate and trans-cinnamaldehyde (LC50 = 44.6 and 43.4 microg/cm2, respectively) while 1,8-cineole showed weak acute toxicity. The essential oil also possessed fumigant toxicity against the booklouse with a LC50 value of 1.5 mg/liter air. 1,8-Cineole and trans-cinnamaldehyde (LC50 = 1.1 and 1.3 mg/liter, respectively) possessed stronger fumigant toxicity against the booklouse than ethyl cinnamate and ethyl rho-methoxycinnamate (LC50 = 10.2 and 10.2 mg/liter air, respectively). trans-Cinnamaldehyde was strongly repellent to booklice, whereas ethyl cinnamate and ethyl rho-methoxycinnamate were weakly repellent and 1,8-cineole did not repel booklice. The results indicate that the essential oil and its constituent compounds have potential for development into natural insecticides or fumigants and repellents for control of insects in stored grains.

The First Report of Tritrichomonas Foetus and Tetratrichomonas Buttreyi in Raccoon Dogs (Nyctereutes Procyonoides) in China.

BACKGROUND: In recent years, the trichomonosis in raccoon dogs in China had occurred frequently. Pentatrichomonas hominis had been described in raccoon dogs in China in some previous studies. PURPOSE TO REVEAL: whether raccoon dogs can be infected by other trichomonad species besides P. hominis, and clarify the prevalence and species distribution of trichomonad in raccoon dogs. METHODS: Herein, the 389 fecal samples were collected from farm-raised raccoon dogs in Hebei Province, all the samples were detected using the microscopic examination and several fecal samples containing trichomonad-like organisms were processed, cultured, stained, and photographed. Meanwhile, all the samples were screened by the species-specific nested PCR based on the small subunit rRNA (SSU rRNA) gene of P. hominis,Tritrichomonas foetus and Tetratrichomonas buttreyi, respectively, and all positive secondary PCR amplications obtained in this study were sequenced, aligned and analysed. RESULTS: 62 fecal samples (15.9%,62/389) were trichomonad-positive under light microscopy, and the trichomonad-like cells were clearly observed in the culture contents. The PCR results showed that 100 samples were trichomonad-positive, including 45 P. hominis-positive samples (11.6%,45/389), 32 T. foetus-positive samples (8.2%,32/389), and 33 T. buttreyi-positive samples (8.5%,33/389), respectively. Double mixed infections were observed in 10 samples. The prevalence of T. foetus and P. hominis were both significantly higher in raccoon dogs with diarrhea (13.9%, and 25.0%) than that in raccoon dogs without diarrhea (7.6%, and 9.3%) (p < 0.05).All samples confirmed as trichomonad-positive under microscopy were also found to be trichomonad-positive by PCR analysis. The sequencing and phylogenetic analysis demonstrated the sequences obtained in this study belonged to P. hominis, T. foetus and T. buttreyi SSU rRNA, respectively. Among them, the T. buttreyi SSU rRNA sequences obtained in this study harbored the new sequence polymorphisms. Based on preliminary morphological and molecular analyses, raccoon dogs are considered as the new host of T. foetus and T. buttreyi. CONCLUSION: This is the first report about the identifcation and prevalence of T. foetus and T. buttreyi in raccoon dogs in China, and the results increase our knowledge about the host range and prevalence of trichomonad species.

Essential oil composition and larvicidal activity of Toddalia asiatica roots against the mosquito Aedes albopictus (Diptera: Culicidae).

The aim of this research was to determine larvicidal activity of the essential oil derived from roots of Toddalia asiatica (L.) Lam. and the isolated constituents against the larvae of the Culicidae mosquito Aedes albopictus. Essential oil of T. asiatica roots was obtained by hydrodistillation and analyzed by GC and GC-MS. A total of 58 components of the essential oil of T. asiatica roots were identified. The essential oil has almost same content of (35.41 %) of sesquiterpenoids and monoterpenoids (31.87 %). The principal compounds in T. asiatica essential oil were geraniol (9.84 %), D-limonene (7.52 %), isopimpinellin (6.62 %), α-gurjunene (6.25 %), and 4-vinylguaiacol (5.94 %). Based on bioactivity-directed fractionation, geraniol, D-limonene, and isopimpinellin were isolated from T. asiatica root essential oil. Geraniol, D-limonene, and isopimpinellin exhibited strong larvicidal activity against A. albopictus with LC(50) values of 30.13, 19.84, and 32.05 µg/ml, respectively, while the essential oil of T. asiatica had an LC(50) value of 69.09 µg/ml. The result indicated that the essential oil of T. asiatica roots and the three isolated constituents have potential for use in control of A. albopictus larvae and could be useful in search of newer, safer, and more effective natural compounds as larvicides.

Identification of insecticidal constituents of the essential oil of Acorus calamus rhizomes against Liposcelis bostrychophila Badonnel.

The aim of this research was to determine the chemical composition of the essential oil of Acorus calamus rhizomes, its insecticidal activity against the booklouse, (Liposcelis bostrychophila) and to isolate any insecticidal constituents from the essential oil. The essential oil of A. calamus rhizomes was obtained by hydrodistillation and analyzed by GC-FID and GC-MS. A total of 32 components of the essential oil of A. calamus rhizomes was identified and the principal compounds in the essential oil were determined to be α-asarone (50.09%), (E)-methylisoeugenol (14.01%), and methyleugenol (8.59%), followed by ß-asarone (3.51%), α-cedrene (3.09%) and camphor (2.42%). Based on bioactivity-guided fractionation, the three active constituents were isolated from the essential oil and identified as methyleugenol, (E)-methylisoeugenol and α-asarone. The essential oil exhibited contact toxicity against L. bostrychophila with an LD50 value of 100.21 µg/cm2 while three constituent compounds, α-asarone, methyleugenol, and (E)-methylisoeugenol had LD50 values of 125.73 µg/cm2, 103.22 µg/cm2 and 55.32 µg/cm2, respectively. Methyleugenol and (E)-methylisoeugenol possessed fumigant toxicity against L. bostrychophila adults with LC50 values of 92.21 µg/L air and 143.43 µg/L air, respectively, while the crude essential oil showed an LC50 value of 392.13 µg/L air. The results indicate that the essential oil of A. calamus rhizomes and its constituent compounds have potential for development into natural fumigants/insecticides for control of the booklice.

Identification of repellent and insecticidal constituents of the essential oil of Artemisia rupestris L. aerial parts against Liposcelis bostrychophila Badonnel.

The aim of this research was to determine the chemical composition and insecticidal and repellent activity of the essential oil of Artemisia rupestris L. aerial parts against the booklice Liposcelis bostrychophila Badonnel and isolation of insecticidal and repellent constituents from the essential oil. The essential oil of A. rupestris was obtained by hydrodistillation and analyzed by GC-MS. A total of 30 components of the essential oil of A. rupestris was identified and the principal compounds in the essential oil were α-terpinyl acetate (37.18%), spathulenol (10.65%), α-terpineol (10.09%), and linalool (7.56%), followed by 4-terpineol (3.92%) and patchoulol (3.05%). Based on bioactivity-guided fractionation, the four active constituents were isolated from the essential oil and identified as α-terpineol, α-terpinyl acetate, 4-terpineol and linalool. The essential oil of A. rupestris exhibited contact toxicity against L. bostrychophila with LD50 value of 414.48 µg/cm². α-Terpinyl acetate (LD50 = 92.59 µg/cm²) exhibited stronger contact toxicity against booklice than α-terpineol (LD50 = 140.30 µg/cm²), 4-terpineol (LD50 = 211.35 µg/cm²), and linalool (LD550 = 393.16 µg/cm²). The essential oil of A. rupestris (LC50 = 6.67 mg/L air) also possessed fumigant toxicity against L. bostrychophila while the four constituents, 4-terpineol, α-terpineol, α-terpinyl acetate and linalool had LC50 values of 0.34, 1.12, 1.26 and 1.96 mg/L air, respectively. α-Terpinol and α-terpinyl acetate showed strong repellency against L. bostrychophila, while linalool and 4-terpinol exhibited weak repellency. The results indicate that the essential oil of A. rupestris aerial parts and its constituent compounds have potential for development into natural insecticides or fumigants as well as repellents for control of insects in stored grains.

Chemical composition and insecticidal activity of the essential oil of Illicium pachyphyllum fruits against two grain storage insects.

The aim of this research was to determine chemical composition and insecticidal activity of the essential oil of Illicium pachyphyllum fruits against two grain storage insects, Sitophilus zeamais and Tribolium castaneum, and to isolate any insecticidal constituents from the essential oil. The essential oil of I. pachyphyllum fruits was obtained by hydrodistillation and analyzed by GC-MS. A total of 36 components of the essential oil were identified, with the principal compounds in the essential oil being trans-ρ-mentha-1(7),8-dien-2-ol (24.56%), D-limonene (9.79%), caryophyllene oxide (9.32%), and cis-carveol (5.26%) followed by ß-caryophyllene (4.63%) and bornyl acetate. Based on bioactivity-guided fractionation, the three active constituents were isolated and identified as trans-ρ-mentha-1(7),8-dien-2-ol, D-limonene and caryophyllene oxide. The essential oil of I. pachyphyllum fruits exhibited contact toxicity against S. zeamais and T. castaneum adults, with LD(50) values of 17.33 µg/adult and 28.94 µg/adult, respectively. trans-p-Mentha-1(7),8-dien-2-ol (LD(50) = 8.66 µg/adult and 13.66 µg/adult, respectively) exhibited stronger acute toxicity against S. zeamais and T. castaneum adults than either caryophyllene oxide (LD(50) = 34.09 µg/adult and 45.56 µg/adult) and D-limonene (LD(50) = 29.86 µg/adult and 20.14 µg/adult). The essential oil of I. pachyphyllum possessed fumigant toxicity against S. zeamais and T. castaneum adults with LC(50) values of 11.49 mg/L and 15.08 mg/L, respectively. trans-p-Mentha-1(7),8-dien-2-ol exhibited stronger fumigant toxicity against S. zeamais and T. castaneum adults, respectively, with LC(50) values of 6.01 mg/L and 8.14 mg/L, than caryophyllene oxide (LC(50) = 17.02 mg/L and 15.98 mg/L) and D-limonene (LC(50) = 33.71 mg/L and 21.24 mg/L). The results indicate that the essential oil of I. pachyphyllum fruits and its constituent compounds have potential for development into natural insecticides or fumigants for the control of insects in stored grains.

First Detection and Molecular Identification of Entamoeba bovis in Farm-Raised Sika Deer from Anhui Province, China.

BACKGROUND: Protozoans of Entamoeba spp. are one of the most common enteric parasites that infect humans and diverse animals including deer. PURPOSE: However, data regarding the prevalence and species/genotypes of Entamoeba spp. in deer in China is scarce. This study investigated the prevalence and species distribution of Entamoeba spp. in sika deer (Cervus nippon) in Anhui Province. METHODS: In our survey, 336 fecal samples were collected from five sika deer farms in different regions of Anhui Province. All samples were examined for the presence of Entamoeba spp. by PCR and phylogenetic analysis of the conserved 18S rRNA gene. RESULTS: 106/336 (31.5%) fecal samples were positive for Entamoeba spp. A statistically significant difference in the prevalence of Entamoeba spp. infection was observed between sampling farms (p < 0.001), and the prevalence of Entamoeba spp. in male and female sika deer showed no significant difference (p > 0.05). Sequence and phylogenetic analysis revealed the single species, E. bovis, was identified in this study. CONCLUSION: This is the first report about the identification of E. bovis in farm-raised sika deer in China, and these results expand our understanding of host range and species distribution of Entamoeba spp. in ruminants.

Detection of Toxoplasma gondii in chicken and soil of chicken farms in Nanjing region, China.

BACKGROUND: Soil is increasingly recognized as an important source in the transmission of Toxoplasma gondii (T. gondii). The aim of this study was to investigate the presence of T. gondii in the soil and to grasp the relationships between the contamination of soil and chicken infections. METHODS: PCR method based on T. gondii-conserved gene internal transcribed spacer 1 (ITS-1) as target gene and ELISA method (sGRA8-ELISA) using the recombinant protein of shortened GRA8 gene of T. gondii as antigen were developed and applied. From April 2013 to March 2014, a total of 700 soil samples were collected at various sites located in thirty farms categorized as free range farm and scale farm in Nanjing, Jiangsu, China, in different seasons. Additionally, a total of 350 sera of chickens were collected from free range farms to determine the presence of antibodies against T. gondii using sGRA8-ELISA. RESULTS: The serological results showed that, antibodies were found in 194 of 250 (67.14%) samples from farms with T. gondii positive in soil and 41 of 100 samples from farms with T. gondii negative in soil (41.00%) (P < 0.01). The PCR detection of soil samples showed that, 7 (2.0%) of 350 samples collected from feeding zone in free range farms were found positive of T. gondii, whereas no sample was positive in scale farms. In the seasonal detections, T. gondii was found in 6 (3.33%) samples collected in autumn and 1 (0.56%) collected in winter. CONCLUSIONS: The results indicated that the contamination of T. gondii in soil in the free range farms was higher than that in the scale farms and seroprevalence of T. gondii in chickens in the farm with soil contamination was higher than that with no soil contamination. The soil contamination might be an effective indicator of T. gondii infection in chickens.

Larvicidal spirobisnaphthalenes from the endophytic fungus Berkleasmium sp. against Aedes albopictus.

BACKGROUND: In our screening programme for new agrochemicals from endophytic fungi, the ethyl acetate extract of an endophytic Berkleasmium sp. isolated from the medicinal plant Dioscorea zingiberensis was found to possess strong larvicidal activity against the Asian tiger mosquito, Aedes albopictus. RESULTS: Bioassay-guided fractionation of the fungal extract has led to the isolation of seven spirobisnaphthalenes, including palmarumycins C8, C12, C15 and B6 and diepoxins γ, δ and ζ. Among them, palmarumycins C8 and B6 showed strong larvicidal activity against the fourth-instar larvae of A. albopictus, with LC50 values of 8.83 and 11.51 µg mL(-1) respectively. Interestingly, only spirobisnaphthalenes with a chlorine substituent possessed strong larvicidal activity. CONCLUSION: The results indicated that the spirobisnaphthalenes derived from the endophytic fungus Berkleasmium sp. could be promising leads for the development of new larvicides against A. albopictus.