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Sample records for cryptocaryon irritans tomont

  1. Ultrastructure observation on the cells at different life history stages of Cryptocaryon irritans (Ciliophora: Prostomatea), a parasitic ciliate of marine fishes.

    PubMed

    Ma, Rui; Ni, Bing; Fan, Xinpeng; Warren, Alan; Yin, Fei; Gu, Fukang

    2016-09-01

    Cells of Cryptocaryon irritans at different life history stages were studied using both light and electron microscopy. The characteristics of several organelles were revealed for the first time at the ultrastructural level. It was confirmed that the cytostome of trophonts, protomonts and theronts was surrounded by cilium-palp triplets rather than ciliary triplets. The nematodesmata underlying the circumoral dikinetids were single bundles, whereas these were always paired in Prorodontids. Toxicysts were present in late-stage tomonts and theronts, but were absent in trophonts and protomonts. We posited that toxicysts might play a role in infection and invasion of host-fish tissue by theronts. The adoral brosse was unlike that of any other family of the class Prostomatea based on its location and morphology. Membranous folds were present in trophonts, protomonts and theronts. These folds were longer and more highly developed in C. irritans than in exclusively free-living prostome ciliates suggesting that they might be linked to parasitism in C. irritans. Trophonts, protomonts and theronts had multiple contractile vacuoles. The basic ultrastructure of the contractile vacuole of C. irritans was similar to that of other kinetofragminophoran ciliates. They might play different roles in different stages of the life cycle since their ultrastructure varied among trophonts, protomonts and theronts. PMID:27460894

  2. Cryptocaryon irritans recombinant proteins as potential antigens for sero-surveillance of cryptocaryonosis.

    PubMed

    Lokanathan, Y; Mohd-Adnan, A; Kua, B-C; Nathan, S

    2016-09-01

    Cryptocaryonosis is a major problem for mariculture, and the absence of suitable sero-surveillance tools for the detection of cryptocaryonosis makes it difficult to screen Cryptocaryon irritans-infected fish, particularly asymptomatic fish. In this study, we proposed a serum-based assay using selected C. irritans proteins to screen infected and asymptomatic fish. Eight highly expressed genes were chosen from an earlier study on C. irritans expressed sequence tags and ciliate glutamine codons were converted to universal glutamine codons. The chemically synthesized C. irritans genes were then expressed in an Escherichia coli expression host under optimized conditions. Five C. irritans proteins were successfully expressed in E. coli and purified by affinity chromatography. These proteins were used as antigens in an enzyme-linked immunosorbent assay (ELISA) to screen sera from experimentally immunized fish and naturally infected fish. Sera from both categories of fish reacted equally well with the expressed C. irritans recombinant proteins as well as with sonicated theronts. This study demonstrated the utility of producing ciliate recombinant proteins in a heterologous expression host. An ELISA was successfully developed to diagnose infected and asymptomatic fish using the recombinant proteins as antigens. PMID:27086498

  3. In vitro treatments for the theront stage of the ciliate protozoan Cryptocaryon irritans.

    PubMed

    Picón-Camacho, S M; Ruiz de Ybáñez, M R; Holzer, A S; Arizcun Arizcun, M; Muñoz, P

    2011-04-01

    The ciliate protozoan Cryptocaryon irritans Brown, 1951, the 'marine white spot', causes one of the most important parasitic fish diseases, with extensive losses every year in mariculture and in the ornamental fish industry. In the present study, we explore the in vitro use of 8 different compounds against the theront (infective) stage of C. irritans; these compounds include extracts of natural products (epigallocatechin gallate (EGCG), L-DOPA, papain), peracetic acid-based compounds (Proxitane 5:23 and 15% peracetic acid, PAA), quinine-based compounds (quinacrine hydrochloride and chloroquine diphosphate) and hydrogen peroxide. All of these compounds had an effect on theront survival; however, only EGCG caused significant theront mortality when applied in doses > or =50 mg l(-1) and over a period of 3 h; papain caused a maximum theront mortality of <50%. We discuss the type of application and potential utility of the compounds tested as part of a management control strategy for C. irritans infections in marine aquaculture and the ornamental fish industry. PMID:21648246

  4. Siganus oramin recombinant L-amino acid oxidase is lethal to Cryptocaryon irritans.

    PubMed

    Li, Ruijun; Dan, Xueming; Li, Anxing

    2013-12-01

    A novel antimicrobial and antiparasitic protein (APP/SR-LAAO) isolated from serum of the rabbitfish (Siganus oramin) was confirmed to be lethal to Cryptocaryon irritans, an important marine parasitic ciliate that causes marine white spot disease in a variety of wild and cultured fish. In this study, a recombinant SR-LAAO (rSR-LAAO) was expressed on a large scale in Escherichia coli Rosetta-gami™(DE3)pLysS cells. rSR-LAAO was expressed as an inactive form in the inclusion bodies. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed that after purification, refolding and ultrafiltration, rSR-LAAO had a significantly cytotoxic effect on C. irritans theronts. Using light microscopy, scanning electron microscopy and fluorescence microscopy, we found that theronts rapidly became weakly motile, cilia became detached, cells became rounded, membranes eventually lysed in different cell positions and cytoplasmic contents leaked out of the cell. These results suggested the recombinant SR-LAAO was significantly lethal to C. irritans and the death process of the parasite incubated with rSR-LAAO was remarkably similar compared to the SR-LAAO group as reported earlier.

  5. A novel protein isolated from the serum of rabbitfish (Siganus oramin) is lethal to Cryptocaryon irritans.

    PubMed

    Wang, Fang-Hua; Xie, Ming-Quan; Li, An-Xing

    2010-07-01

    The susceptibility of eight marine fish species cultured in South China were tested for infection by the parasitic ciliate, Cryptocaryon irritans, via a challenge examination and an immobilization assay. All species of fish (representing six different families) that we investigated were infected by C. irritans except the rabbitfish (Siganus oramin), which displayed resistance to C. irritans infection. The infection intensity of rabbitfish (0.92+/-0.97, p<0.05) was significantly lower while the immobilization titres of rabbitfish serum were significantly higher (44.51+/-22.98, p<0.05) than the other seven species of fish. Additionally, the serum of the rabbitfish presented a strong killing effect to C. irritans in vitro. Light microscopy, scanning electron microscopy and fluorescence microscopy confirmed that rabbitfish serum could induce the theront cilia fall off, rupture of the cell membrane because of the swell and rupture of the macronucleus. Rabbitfish serum could also induce the rupture of the trophont membrane and content efflux. Herein a novel antiparasitic protein (APP) was isolated and purified from the serum of rabbitfish (S. oramin) by using a series of salting-out, cation exchange chromatography and two step of reversed phase high performance liquid chromatography (RP-HPLC). Analysis of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed that APP was a homogenous polymeric protein with an N-terminal amino acid sequence of SSVEKNLAACLRDND. Its monomeric molecular mass, determined by matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometer (MALDI-TOF-TOF-MS), was found to be 61,739.87 Da. Results of homology analyses indicated that this protein was a newly discovered functional protein in the rabbitfish serum. Laser confocal fluorescence microscopy conformed that the action site of the APP was mainly on the cell membrane and nucleus of theront, which agreed with the results of light microscopy

  6. Effects of Cryptocaryon irritans infection on the survival, feeding, respiratory rate and ionic regulation of the marbled rockfish Sebastiscus marmoratus.

    PubMed

    Yin, Fei; Gong, Qiyang; Li, Yanwei; Dan, Xueming; Sun, Peng; Gao, Quanxin; Shi, Zhaohong; Peng, Shiming; Li, Anxing

    2014-02-01

    To clarify the effects of a Cryptocaryon irritans infection on the physiological functions of the marbled rockfish Sebastiscus marmoratus, this study utilized C. irritans at concentrations of 2500; 5000; 7500; 10,000; 20,000; and 30,000 theronts/fish to infect marbled rockfish weighing 45 ± 3 g. The survival rate, food intake, respiratory rate, serum ion concentrations and gill Na+/K+-ATPase activity were determined. With the increase of the infection concentration and the passage of time, the survival rate of the rockfish gradually decreased. The groups infected with more than 5000 theronts/fish had stopped feeding within 4 days. The respiratory rates of the fish in the groups infected with 2500 and 5000 theronts/fish initially increased and then decreased. In contrast, the respiratory rate of the fish in the groups infected with more than 7500 theronts/fish was elevated to levels significantly higher than the control group after 12 h. The Na+/K+-ATPase activity and serum Na+ and Cl- concentrations increased with increasing infection concentration. In conclusion, the physiological functions of the fish infected with low concentrations of C. irritans can be effectively restored, whereas a high concentration infection induced severe stress. The declined food intake and accelerated respiratory rate could be useful for an early warning system as important indicators.

  7. Grouper (Epinephelus coioides) BCR signaling pathway was involved in response against Cryptocaryon irritans infection.

    PubMed

    Mo, Ze-Quan; Yang, Man; Wang, Hai-Qing; Xu, Yang; Huang, Mian-Zhi; Lao, Guo-Feng; Li, Yan-Wei; Li, An-Xing; Luo, Xiao-Chun; Dan, Xue-Ming

    2016-10-01

    B cell antigen receptor (BCR) plays a crucial role in B cell development and antibody production. It comprises membrane immunoglobulin non-covalently associated with CD79a/CD79b heterodimer. After B cell activation, initial extracellular signals are transduced by BCR complex and amplified by two protein tyrosine kinases, LYN and SYK, which then trigger various pathways. In the present study, we cloned grouper genes for BCR accessory molecules, EcCD79a (669 bp) and EcCD79b (639 bp), as well as two protein tyrosine kinases, EcLYN (1482 bp) and EcSYK (1854 bp). Homology analysis showed that all four molecules had a relatively high amino acid identity compared with those in other animals. Among them, they all shared the highest identity with Takifugu rubripes (EcCD79a 49%, EcCD79b 52%, EcLYN 82% and EcSYK 77%). The conserved features and important functional residues were analyzed. Together with IgM and IgT, tissue distribution analysis showed that all six molecules were mainly expressed in immune organs, particularly systematic immune organs. In groupers infected with Cryptocaryon irritans, up-regulation of EcCD79a and b, EcIgM and EcIgT were not seen in the early stage skin and gill until 14-21 days. Up-regulation of EcCD79a was seen in head kidney at most time points, while EcCD79a and b were only significantly up-regulated in day 14 spleen. Significant up-regulation of EcIgT were seen in day 21 head kidney and day 1, day14 spleen. Significant up-regulation of EcIgM were seen in day 1 head kidney and 12 h spleen. In addition, two protein kinase genes, EcLYN and EcSYK, were up-regulated in the skin at most time points, which suggested that B cells may be activated at the skin local infection site. PMID:27514788

  8. Immune responses and immune-related gene expression profile in orange-spotted grouper after immunization with Cryptocaryon irritans vaccine.

    PubMed

    Dan, Xue-Ming; Zhang, Tuan-Wei; Li, Yan-Wei; Li, An-Xing

    2013-03-01

    In order to elucidate the immune-protective mechanisms of inactivated Cryptocaryon irritans vaccine, different doses of C. irritans theronts were used to immunize orange-spotted grouper (Epinephelus coioides). We measured serum immobilization titer, blood leukocyte respiratory burst activity, serum alternative complement activity, and serum lysozyme activity weekly. In addition, the expression levels of immune-related genes such as interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), major histocompatibility complexes I and II (MHC I and II), and transforming growth factor-β1 (TGF-β1) were determined in spleen and gills. The results showed that the immobilization titer, respiratory burst activity, and alternative complement activity of immunized fish were significantly increased, and the levels of the last two immune parameters in the high-dose vaccine group were significantly higher than in the low-dose vaccine group. Serum lysozyme activity in the high-dose vaccine group was significantly higher than in the PBS control group. Vaccination also regulated host immune-related gene expression. For example, at 2- and 3- weeks post immunization, IL-1β expression in the high-dose vaccine group spleen was significantly increased. At 4-weeks post immunization, the fish were challenged with a lethal dose of parasite, and the survival rates of high-dose vaccine group, low-dose vaccine group, PBS control group, and adjuvant control group were 80%, 40%, 0%, and 10% respectively. These results demonstrate that inactivated C. irritans vaccination improves specific and nonspecific immune responses in fish, enhancing their anti-parasite ability. These effects are vaccine antigen dose-dependent.

  9. Molecular cloning of NCCRP-1 gene from orange-spotted grouper (Epinephelus coioides) and characterization of NCCRP-1(+) cells post Cryptocaryon irritans infection.

    PubMed

    Huang, Xia-Zi; Li, Yan-Wei; Mai, Yong-Zhan; Luo, Xiao-Chun; Dan, Xue-Ming; Li, An-Xing

    2014-10-01

    Nonspecific cytotoxic cells (NCCs) are an important cytotoxic cell population in the innate teleost immune system. The receptor designated "NCC receptor protein 1" (NCCRP-1) has been reported to be involved in the recognition and activation of NCCs. In this study, the full-length cDNA of Epinephelus coioides NCCRP-1 (ecnccrp-1) was cloned. The open reading frame (ORF) of ecnccrp-1 is 699 bp, encoding a 232 amino acid protein that includes proline-rich motifs at the N-terminus and is related to the F-box associated family. Although a bioinformatics analysis showed that EcNCCRP-1 had no signal peptide or transmembrane helices, a polyclonal antibody directed against recombinant EcNCCRP-1 efficiently labeled a membrane protein in the head kidney, detected with Western blot analysis, which indicated that the protein localized to the cell surface. RT-PCR showed that the constitutive expression of ecnccrp-1 was higher in the lymphoid organs, such as the trunk kidney, spleen, head kidney, and thymus, and lower in brain, heart, fat, liver, muscle, and skin. After infection with Cryptocaryon irritans, the transcription of ecnccrp-1 was analyzed at the infected sites (skin and gills) and in the systemic immune organs (head kidney and spleen). At the infected sites, especially the skin, ecnccrp-1 expression was upregulated at 6h post infection, reaching peak expression on day 3 post the primary infection. However, the expression patterns differed in the systemic immune organs. In the spleen, ecnccrp-1 was gradually increased in the early infection period and decreased sharply on day 3 post the primary infection, whereas in the head kidney, the transcription of ecnccrp-1 was depressed during almost the whole course of infection. An immunohistochemical analysis showed that EcNCCRP-1(+) cells accumulated at the sites of infection with C. irritans. These results suggested that NCCs were involved in the process of C. irritans infection in E. coioides.

  10. Molecular cloning of NCCRP-1 gene from orange-spotted grouper (Epinephelus coioides) and characterization of NCCRP-1(+) cells post Cryptocaryon irritans infection.

    PubMed

    Huang, Xia-Zi; Li, Yan-Wei; Mai, Yong-Zhan; Luo, Xiao-Chun; Dan, Xue-Ming; Li, An-Xing

    2014-10-01

    Nonspecific cytotoxic cells (NCCs) are an important cytotoxic cell population in the innate teleost immune system. The receptor designated "NCC receptor protein 1" (NCCRP-1) has been reported to be involved in the recognition and activation of NCCs. In this study, the full-length cDNA of Epinephelus coioides NCCRP-1 (ecnccrp-1) was cloned. The open reading frame (ORF) of ecnccrp-1 is 699 bp, encoding a 232 amino acid protein that includes proline-rich motifs at the N-terminus and is related to the F-box associated family. Although a bioinformatics analysis showed that EcNCCRP-1 had no signal peptide or transmembrane helices, a polyclonal antibody directed against recombinant EcNCCRP-1 efficiently labeled a membrane protein in the head kidney, detected with Western blot analysis, which indicated that the protein localized to the cell surface. RT-PCR showed that the constitutive expression of ecnccrp-1 was higher in the lymphoid organs, such as the trunk kidney, spleen, head kidney, and thymus, and lower in brain, heart, fat, liver, muscle, and skin. After infection with Cryptocaryon irritans, the transcription of ecnccrp-1 was analyzed at the infected sites (skin and gills) and in the systemic immune organs (head kidney and spleen). At the infected sites, especially the skin, ecnccrp-1 expression was upregulated at 6h post infection, reaching peak expression on day 3 post the primary infection. However, the expression patterns differed in the systemic immune organs. In the spleen, ecnccrp-1 was gradually increased in the early infection period and decreased sharply on day 3 post the primary infection, whereas in the head kidney, the transcription of ecnccrp-1 was depressed during almost the whole course of infection. An immunohistochemical analysis showed that EcNCCRP-1(+) cells accumulated at the sites of infection with C. irritans. These results suggested that NCCs were involved in the process of C. irritans infection in E. coioides. PMID:24844613

  11. Immunological, ionic and biochemical responses in blood serum of the marine fish Trachinotus ovatus to poly-infection by Cryptocaryon irritans.

    PubMed

    Yin, Fei; Sun, Peng; Tang, Baojun; Dan, Xueming; Li, Anxing

    2015-07-01

    To investigate the response of pompano fish (Trachinotus ovatus) to white spot disease, we used the protozoan Cryptocaryon irritans to infect live 450-g specimens at concentrations of 40,000 theronts/fish. We assessed the relative infection intensity (RII), serum immobilizing titer, and immunity-related enzyme activities (ACP, AKP, LZM), and assessed feeding, serum ion concentrations (Na(+), Cl(-), Ca(2+) and K(+)) and blood biochemistry (ALT, AST, LDH) of pompano. The fish were then treated with a lethal dose of C. irritans (70,000 theronts/fish) and the number of deaths was recorded. We found that the relative infection intensities of the control group, group I, and group II were 0, 0.630 ± 0.179, and 0.014 ± 0.006. Poly-infection induced a significant increase in the serum immobilizing titer (853.33 ± 295.60) of group II. In terms of the biochemical assessment, group II had significantly higher alkaline phosphatase and acid phosphatase activities than the other groups, and the lowest lysozyme activity (P < 0.05), compared to higher activity in the control group and the highest level in group I. Only the fishes of group I had stopped feeding after treatment. The concentrations of Na(+), Cl(-), and Ca(2+) in blood serum did not differ significantly among the three groups, but K(+) concentration increased with the increasing infection frequency. Alanine aminotransferase, aspartate aminotransferase, and lactate dehydrogenase activities in fish of group II were significantly higher than those of the other groups. Survival of the fish subjected to the lethal dose of C. irritans was 0, 0, and 100 in groups control, I, and II, respectively. In conclusions, based on the food intake of group II, along with the results of relative infection intensity, serum immobilizing titer, and survival, we speculate that the fish in that group acquired high protective immunity following poly-infection by C. irritans, experiencing limited harm for pompano.

  12. Characterization of E3 ubiquitin ligase neuregulin receptor degradation protein-1 (Nrdp1) in the large yellow croaker (Larimichthys crocea) and its immune responses to Cryptocaryon irritans.

    PubMed

    Zhang, Dong Ling; Han, Fang; Yu, Da Hui; Xiao, Shi Jun; Li, Ming Yun; Chen, Jian; Wang, Zhi Yong

    2015-02-10

    Neuregulin receptor degradation protein-1 (Nrdp1) was recently identified in humans as an important immune factor responding to the challenge of virus, LPS or cytokine. Its role in fish immune defense and whether it is involved in anti-parasite immunity have not been proven yet. In this report, the full-length cDNA sequence and genomic structure of Nrdp1 in the large yellow croaker Larimichthys crocea (LcNrdp1) were identified and characterized. The full-length cDNA of LcNrdp1 was 1248bp, including a 5' untranslated region (UTR) of 32bp, a 3' UTR of 259bp and an open reading frame (ORF) of 937bp, encoding a polypeptide of 318 amino acid residues. The full-length genomic DNA sequence of LcNrdp1 was composed of 2635 nucleotides, including four exons and three introns. The putative LcNrdp1 protein had no signal peptide sequence and contained a characteristic Nrdp1 consensus motif C3HC3D ring finger and a Coiled-coil domain. Phylogenetic analysis showed that Nrdp1 in fish was closer with that in other vertebrates (79%-90% amino acid identity) than in invertebrates and bacteria (27%-65%). In fishes, Nrdp1 in large yellow croaker was closer with that in Takifugu rubripes. The expression profile showed that LcNrdp1 was constitutively expressed in all tested tissues, especially highly expressed in brain, muscle and kidney. Post-infection (PI) with Cryptocaryon irritans, an increased expression of LcNrdp1 was induced in infection sites (skin and gill), whereas in immune organs, the expression of LcNrdp1 was up-regulated in spleen (except the 1st d and 10th d PI) but suppressed in head kidney. These results suggested that LcNrdp1 might play an important immune role in the finfish L. crocea in the defense against the parasite C. irritans. PMID:25447921

  13. Characterization of E3 ubiquitin ligase neuregulin receptor degradation protein-1 (Nrdp1) in the large yellow croaker (Larimichthys crocea) and its immune responses to Cryptocaryon irritans.

    PubMed

    Zhang, Dong Ling; Han, Fang; Yu, Da Hui; Xiao, Shi Jun; Li, Ming Yun; Chen, Jian; Wang, Zhi Yong

    2015-02-10

    Neuregulin receptor degradation protein-1 (Nrdp1) was recently identified in humans as an important immune factor responding to the challenge of virus, LPS or cytokine. Its role in fish immune defense and whether it is involved in anti-parasite immunity have not been proven yet. In this report, the full-length cDNA sequence and genomic structure of Nrdp1 in the large yellow croaker Larimichthys crocea (LcNrdp1) were identified and characterized. The full-length cDNA of LcNrdp1 was 1248bp, including a 5' untranslated region (UTR) of 32bp, a 3' UTR of 259bp and an open reading frame (ORF) of 937bp, encoding a polypeptide of 318 amino acid residues. The full-length genomic DNA sequence of LcNrdp1 was composed of 2635 nucleotides, including four exons and three introns. The putative LcNrdp1 protein had no signal peptide sequence and contained a characteristic Nrdp1 consensus motif C3HC3D ring finger and a Coiled-coil domain. Phylogenetic analysis showed that Nrdp1 in fish was closer with that in other vertebrates (79%-90% amino acid identity) than in invertebrates and bacteria (27%-65%). In fishes, Nrdp1 in large yellow croaker was closer with that in Takifugu rubripes. The expression profile showed that LcNrdp1 was constitutively expressed in all tested tissues, especially highly expressed in brain, muscle and kidney. Post-infection (PI) with Cryptocaryon irritans, an increased expression of LcNrdp1 was induced in infection sites (skin and gill), whereas in immune organs, the expression of LcNrdp1 was up-regulated in spleen (except the 1st d and 10th d PI) but suppressed in head kidney. These results suggested that LcNrdp1 might play an important immune role in the finfish L. crocea in the defense against the parasite C. irritans.

  14. Co-expression of march5b and tlr7 in large yellow croaker Larimichthys crocea in response to Cryptocaryon irritans infection.

    PubMed

    Zhang, D L; Yu, D H; Chen, J; Chen, C; Wang, Z Y

    2015-08-01

    In this study, molecular characteristics of march5b and co-expression of march5b and tlr7 in response to the infection of Cryptocaryon irritans in the large yellow croaker Larimichthys crocea were investigated. The full-length complementary (c)DNA of march5b was 1314 bp, including an open reading frame of 846 bp encoding a polypeptide of 281 amino acids, and the full-length genomic sequence was composed of 23,577 nucleotides, including six exons and five introns. The putative March5b protein contained a RINGv motif and four transmembrane domains. The march5b transcripts were broadly distributed in all detected tissues, with a strong expression in blood, brain and gills, and a weak expression in kidney by quantitative PCR analysis. The expression of march5b and tlr7 in the skin, gills, spleen and head kidney changed in the same manner at most time points post-primary infection with C. irritans. Significant increase was observed in the skin with march5b at days 2 and 3 by 26.10 and 6.88 fold, respectively, and with tlr7 at day 3 by 57.68 fold, when compared with the control. Their expressions, however, were decreased in the gills, especially at day 3 (march5b by 8.9%, tlr7 by 22.06%). In the spleen and head kidney, march5b and tlr7 transcripts were up-regulated early, then noticeably declined at day 3. These results suggested that march5b and tlr7 are co-expressed in response to parasite infection and March5b probably catalyses ubiquitination of some proteins of TLR7 signalling pathway.

  15. Growth, feed intake and immune responses of orange-spotted grouper (Epinephelus coioides) exposed to low infectious doses of ectoparasite (Cryptocaryon irritans).

    PubMed

    Yin, Fei; Dan, Xue-Ming; Sun, Peng; Shi, Zhao-Hong; Gao, Quan-Xin; Peng, Shi-Ming; Li, An-Xing

    2014-01-01

    To explore the effect of low-dose Cryptocaryon irritans infection on growth, feeding and antiparasitic immunity of orange-spotted grouper (Epinephelus coioides), this study utilized C. irritans at concentrations of 5500 theronts/fish (Group I, 1/10 of 96 h LC50) or 11,000 theronts/fish (Group II) to infect E. coioides weighing 38 g on average at week 0, 2 and 4, respectively. Food consumption was recorded daily; the fish were weighed weekly; serum immobilizing titer (SIT), and acid phosphatase (ACP), alkaline phosphatase (AKP), superoxide dismutase (SOD), lysozyme (LZM) activity were recorded every 2 weeks; the fish were treated with lethal dose (70,000 theronts/fish) of C. irritans in the 8th week and death number were recorded. The result shows that in the 1st week after the first infection, the fish's weight gain (WG), length gain (LG), and specific growth rate (SGR) dropped as parasite dose increased, and WG, SGR values were negative; while, after the 2nd and the 3rd infection, no significant differences were detected among the three groups. These results indicated that the 1st infection affected the fish most, while the following infections were protected by some immunity. In the 3rd, 7th, and 8th week, condition factor (CF) increased with the increased infectious dose, indicating that the parasite affected body length more than body weight. As the experiment went on, accumulated food consumption (AFC) of all three groups steadily grew (control > Group I > Group II). But on the 2nd day after the first infection, daily food consumption (DFC) of Group I and II significantly dropped, the decline of Group II was greater than that of Group I, DFC recovered in the following week, with Group I earlier than Group II. After the 2nd infection, DFC of Group I and II dropped again, Group II still dropped more than Group I, and both groups recovered on the 3rd day after infection. The 3rd infection caused no significant difference in week food consumption (WFC). These

  16. The Rab5A gene of marine fish, large yellow croaker (Larimichthys crocea), and its response to the infection of Cryptocaryon irritans.

    PubMed

    Han, Fang; Zhang, Yu; Zhang, Dongling; Liu, Lanping; Tsai, Huai Jen; Wang, Zhiyong

    2016-07-01

    Rab GTPases, members of the Ras superfamily, encode monomeric G-proteins. Rab proteins regulate key steps in membrane traffic transport and endocytic pathway of host immune responses. Rab5A is involved in immune regulation, particularly in T cell migration and macrophage endocytosis in higher vertebrates. However, little is known of the molecular structure of Rab5A gene in marine teleost fish species and its expression profile during the parasite infection. In this study, the full-length cDNA sequence and genomic structure of Rab5A gene of the large yellow croaker (Larimichthys crocea) (LycRab5A), one of the most economical marine fishes, were identified and characterized. The LycRab5A protein, containing the ATPase/GTPase binding motifs and the effector molecules binding motifs, was highly homologous to that of other animals. The expression plasmid containing LycRab5A cDNA fused with GST was engineered and transformed into Escherichia coli to produce recombinant protein GST-LycRab5A, which was purified to prepare a polyclonal antibody specifically against LycRab5A. Subcellular localization revealed that LycRab5A expressed in the membrane and cytoplasm. Based on real-time PCR and Western blot analysis, we found that both mRNA and protein of LycRab5A were expressed in all tissues we examined; especially it was highly expressed in blood and gill. Interestingly, both mRNA and protein of LycRab5A were substantially up-regulated when parasitic ciliate protozoan (Cryptocaryon irritans) was infected. The expression of LycRab5A was reached to the maximal level at 24 h after infection. The line of evidence suggested that LycRab5A might play an important role in large yellow croaker defense against parasite infection. Moreover, on the basis of protein interaction, it was found that the LycRab5A interacted with myosin light chain (designated as LycMLC), a crucial protein in the process of phagocytosis. This discovery might contribute better understanding to the molecular

  17. Detecting Burrowing Owl Bloodmeals in Pulex irritans (Siphonaptera: Pulicidae).

    PubMed

    Graham, Christine B; Eisen, Rebecca J; Belthoff, James R

    2016-03-01

    Pulex irritans L. is a cosmopolitan flea species that infests a wide variety of hosts. In North America it generally parasitizes large wild mammals, but in the Pacific Northwest an association has emerged between P. irritans and the western burrowing owl (Athene cunicularia hypugaea). While investigators have recognized this association for decades, it has not been clear if P. irritans feeds on burrowing owls, or if the owls serve exclusively as phoretic hosts. Here we describe using a real-time assay that was originally developed to identify bloodmeals in Ugandan cat fleas (Ctenocephalides felis Bouché) to detect burrowing owl DNA in P. irritans collected from burrowing owls in southern Idaho. Of 50 fleas tested, 12 had no detectable vertebrate bloodmeal. The remaining 38 (76%) contained burrowing owl DNA. The assay did not detect vertebrate DNA in unfed fleas exposed to owl or mouse pelts and is therefore unlikely to detect DNA in fleas from vertebrates that have served exclusively as phoretic hosts. We conclude that P. irritans feeds on burrowing owls. We discuss the potential implications of this finding for burrowing owl conservation and enzootic plague dynamics.

  18. Detecting Burrowing Owl Bloodmeals in Pulex irritans (Siphonaptera: Pulicidae).

    PubMed

    Graham, Christine B; Eisen, Rebecca J; Belthoff, James R

    2016-03-01

    Pulex irritans L. is a cosmopolitan flea species that infests a wide variety of hosts. In North America it generally parasitizes large wild mammals, but in the Pacific Northwest an association has emerged between P. irritans and the western burrowing owl (Athene cunicularia hypugaea). While investigators have recognized this association for decades, it has not been clear if P. irritans feeds on burrowing owls, or if the owls serve exclusively as phoretic hosts. Here we describe using a real-time assay that was originally developed to identify bloodmeals in Ugandan cat fleas (Ctenocephalides felis Bouché) to detect burrowing owl DNA in P. irritans collected from burrowing owls in southern Idaho. Of 50 fleas tested, 12 had no detectable vertebrate bloodmeal. The remaining 38 (76%) contained burrowing owl DNA. The assay did not detect vertebrate DNA in unfed fleas exposed to owl or mouse pelts and is therefore unlikely to detect DNA in fleas from vertebrates that have served exclusively as phoretic hosts. We conclude that P. irritans feeds on burrowing owls. We discuss the potential implications of this finding for burrowing owl conservation and enzootic plague dynamics. PMID:26545716

  19. Burrowing Owls, Pulex irritans, and Plague.

    PubMed

    Belthoff, James R; Bernhardt, Scott A; Ball, Christopher L; Gregg, Michael; Johnson, David H; Ketterling, Rachel; Price, Emily; Tinker, Juliette K

    2015-09-01

    Western Burrowing Owls (Athene cunicularia hypugaea) are small, ground-dwelling owls of western North America that frequent prairie dog (Cynomys spp.) towns and other grasslands. Because they rely on rodent prey and occupy burrows once or concurrently inhabited by fossorial mammals, the owls often harbor fleas. We examined the potential role of fleas found on burrowing owls in plague dynamics by evaluating prevalence of Yersinia pestis in fleas collected from burrowing owls and in owl blood. During 2012-2013, fleas and blood were collected from burrowing owls in portions of five states with endemic plague-Idaho, Oregon, Washington, Colorado, and South Dakota. Fleas were enumerated, taxonomically identified, pooled by nest, and assayed for Y. pestis using culturing and molecular (PCR) approaches. Owl blood underwent serological analysis for plague antibodies and nested PCR for detection of Y. pestis. Of more than 4750 fleas collected from owls, Pulex irritans, a known plague vector in portions of its range, comprised more than 99.4%. However, diagnostic tests for Y. pestis of flea pools (culturing and PCR) and owl blood (PCR and serology) were negative. Thus, even though fleas were prevalent on burrowing owls and the potential for a relationship with burrowing owls as a phoretic host of infected fleas exists, we found no evidence of Y. pestis in sampled fleas or in owls that harbored them. We suggest that studies similar to those reported here during plague epizootics will be especially useful for confirming these results. PMID:26367482

  20. Burrowing Owls, Pulex irritans, and Plague.

    PubMed

    Belthoff, James R; Bernhardt, Scott A; Ball, Christopher L; Gregg, Michael; Johnson, David H; Ketterling, Rachel; Price, Emily; Tinker, Juliette K

    2015-09-01

    Western Burrowing Owls (Athene cunicularia hypugaea) are small, ground-dwelling owls of western North America that frequent prairie dog (Cynomys spp.) towns and other grasslands. Because they rely on rodent prey and occupy burrows once or concurrently inhabited by fossorial mammals, the owls often harbor fleas. We examined the potential role of fleas found on burrowing owls in plague dynamics by evaluating prevalence of Yersinia pestis in fleas collected from burrowing owls and in owl blood. During 2012-2013, fleas and blood were collected from burrowing owls in portions of five states with endemic plague-Idaho, Oregon, Washington, Colorado, and South Dakota. Fleas were enumerated, taxonomically identified, pooled by nest, and assayed for Y. pestis using culturing and molecular (PCR) approaches. Owl blood underwent serological analysis for plague antibodies and nested PCR for detection of Y. pestis. Of more than 4750 fleas collected from owls, Pulex irritans, a known plague vector in portions of its range, comprised more than 99.4%. However, diagnostic tests for Y. pestis of flea pools (culturing and PCR) and owl blood (PCR and serology) were negative. Thus, even though fleas were prevalent on burrowing owls and the potential for a relationship with burrowing owls as a phoretic host of infected fleas exists, we found no evidence of Y. pestis in sampled fleas or in owls that harbored them. We suggest that studies similar to those reported here during plague epizootics will be especially useful for confirming these results.

  1. Simultaneous detection of Pyrethroid, Organophosphate and Cyclodiene target site resistance in Haematobia irritans (Diptera: Muscidae) by multiplex Polymerase chain reaction

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The horn fly, Haematobia irritans irritans (Linnaeus, 1758), is an important pest that causes significant economic losses to the livestock industry, but insecticide resistance in horn fly populations has made horn fly control increasingly difficult to achieve. In this study, we developed a multiplex...

  2. Wolbachia infection in Australasian and North American populations of Haematobia irritans (Diptera: Muscidae).

    PubMed

    Zhang, Bing; McGraw, Elizabeth; Floate, Kevin D; James, Peter; Jorgensen, Wayne; Rothwell, Jim

    2009-06-10

    Buffalo fly (Haematobia irritans exigua) is a major pest of beef and dairy cattle in northern Australia. Global warming is expected to increase the southern range of buffalo fly. Chemical control is compromised by resistance and may not be feasible in extensive production systems and there is rapidly growing market preference for beef and dairy products produced in low-chemical systems. Wolbachia are vertically transmitted intracellular bacteria that can profoundly influence host reproduction and fitness and are of increasing interest for use in biocontrol programs. To determine whether Australian flies are infected with Wolbachia, buffalo flies were collected from 12 cattle herds around Australia and assayed by standard PCR for the Wolbachia wsp gene. H. i. exigua from Indonesia and horn fly (H. i. irritans) from Canada were also tested. All H. i. exigua samples tested were negative for Wolbachia infection whereas a very strong signal for Wolbachia was obtained from H. i. irritans.

  3. Functional genomics of the horn fly, Haematobia irritans (Linnaeus, 1758)

    PubMed Central

    2011-01-01

    Background The horn fly, Haematobia irritans (Linnaeus, 1758) (Diptera: Muscidae) is one of the most important ectoparasites of pastured cattle. Horn flies infestations reduce cattle weight gain and milk production. Additionally, horn flies are mechanical vectors of different pathogens that cause disease in cattle. The aim of this study was to conduct a functional genomics study in female horn flies using Expressed Sequence Tags (EST) analysis and RNA interference (RNAi). Results A cDNA library was made from whole abdominal tissues collected from partially fed adult female horn flies. High quality horn fly ESTs (2,160) were sequenced and assembled into 992 unigenes (178 contigs and 814 singlets) representing molecular functions such as serine proteases, cell metabolism, mitochondrial function, transcription and translation, transport, chromatin structure, vitellogenesis, cytoskeleton, DNA replication, cell response to stress and infection, cell proliferation and cell-cell interactions, intracellular trafficking and secretion, and development. Functional analyses were conducted using RNAi for the first time in horn flies. Gene knockdown by RNAi resulted in higher horn fly mortality (protease inhibitor functional group), reduced oviposition (vitellogenin, ferritin and vATPase groups) or both (immune response and 5'-NUC groups) when compared to controls. Silencing of ubiquitination ESTs did not affect horn fly mortality and ovisposition while gene knockdown in the ferritin and vATPse functional groups reduced mortality when compared to controls. Conclusions These results advanced the molecular characterization of this important ectoparasite and suggested candidate protective antigens for the development of vaccines for the control of horn fly infestations. PMID:21310032

  4. Rdl-containing Fragment of GABA(A) from the Horn Fly, Haematobia Irritans, Susceptible Genotype

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The horn fly, Haematobia irritans (L.), is a significant economic pest of cattle found throughout Europe, North Africa, Asia Minor, and the Americas. The major means of controlling the horn fly is through applications of chemicals with insecticidal activity. A cyclodiene-containing ear tag product h...

  5. 454 pyrosequencing project identifying expressed genes from the horn fly, Haematobia irritans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We used an EST approach to initiate a study of the genome of the horn fly, Haematobia irritans and have used 454 pyrosequencing techniques to sequence 73,512, 100,603, 71,550, and 85,769 expressed genes from the egg, first instar larvae, adult male, and adult female lifestages of the horn fly. cD...

  6. Acetylcholinesterase of Haematobia irritans (Diptera: Muscidae): Baculovirus expression, biochemical properties and organophosphate insensitivity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study reports the baculovirus expression and biochemical characterization of recombinant acetylcholinesterase from Haematobia irritans (L) (rHiAChE) and the effect of the previously described G262A mutation on enzyme activity and sensitivity to selected organophosphates. The rHiAChE was confirm...

  7. Characterization of irritans mariner-like elements in the olive fruit fly Bactrocera oleae (Diptera: Tephritidae): evolutionary implications.

    PubMed

    Ben Lazhar-Ajroud, Wafa; Caruso, Aurore; Mezghani, Maha; Bouallegue, Maryem; Tastard, Emmanuelle; Denis, Françoise; Rouault, Jacques-Deric; Makni, Hanem; Capy, Pierre; Chénais, Benoît; Makni, Mohamed; Casse, Nathalie

    2016-08-01

    Genomic variation among species is commonly driven by transposable element (TE) invasion; thus, the pattern of TEs in a genome allows drawing an evolutionary history of the studied species. This paper reports in vitro and in silico detection and characterization of irritans mariner-like elements (MLEs) in the genome and transcriptome of Bactrocera oleae (Rossi) (Diptera: Tephritidae). Eleven irritans MLE sequences have been isolated in vitro using terminal inverted repeats (TIRs) as primers, and 215 have been extracted in silico from the sequenced genome of B. oleae. Additionally, the sequenced genomes of Bactrocera tryoni (Froggatt) and Bactrocera cucurbitae (Diptera: Tephritidae) have been explored to identify irritans MLEs. A total of 129 sequences from B. tryoni have been extracted, while the genome of B. cucurbitae appears probably devoid of irritans MLEs. All detected irritans MLEs are defective due to several mutations and are clustered together in a monophyletic group suggesting a common ancestor. The evolutionary history and dynamics of these TEs are discussed in relation with the phylogenetic distribution of their hosts. The knowledge on the structure, distribution, dynamic, and evolution of irritans MLEs in Bactrocera species contributes to the understanding of both their evolutionary history and the invasion history of their hosts. This could also be the basis for genetic control strategies using transposable elements. PMID:27392643

  8. Characterization of irritans mariner-like elements in the olive fruit fly Bactrocera oleae (Diptera: Tephritidae): evolutionary implications

    NASA Astrophysics Data System (ADS)

    Ben Lazhar-Ajroud, Wafa; Caruso, Aurore; Mezghani, Maha; Bouallegue, Maryem; Tastard, Emmanuelle; Denis, Françoise; Rouault, Jacques-Deric; Makni, Hanem; Capy, Pierre; Chénais, Benoît; Makni, Mohamed; Casse, Nathalie

    2016-08-01

    Genomic variation among species is commonly driven by transposable element (TE) invasion; thus, the pattern of TEs in a genome allows drawing an evolutionary history of the studied species. This paper reports in vitro and in silico detection and characterization of irritans mariner-like elements (MLEs) in the genome and transcriptome of Bactrocera oleae (Rossi) (Diptera: Tephritidae). Eleven irritans MLE sequences have been isolated in vitro using terminal inverted repeats (TIRs) as primers, and 215 have been extracted in silico from the sequenced genome of B. oleae. Additionally, the sequenced genomes of Bactrocera tryoni (Froggatt) and Bactrocera cucurbitae (Diptera: Tephritidae) have been explored to identify irritans MLEs. A total of 129 sequences from B. tryoni have been extracted, while the genome of B. cucurbitae appears probably devoid of irritans MLEs. All detected irritans MLEs are defective due to several mutations and are clustered together in a monophyletic group suggesting a common ancestor. The evolutionary history and dynamics of these TEs are discussed in relation with the phylogenetic distribution of their hosts. The knowledge on the structure, distribution, dynamic, and evolution of irritans MLEs in Bactrocera species contributes to the understanding of both their evolutionary history and the invasion history of their hosts. This could also be the basis for genetic control strategies using transposable elements.

  9. High chromosomal variation in wild horn fly Haematobia irritans (Linnaeus) (Diptera, Muscidae) populations

    PubMed Central

    Forneris, Natalia S.; Otero, Gabriel; Pereyra, Ana; Repetto, Gustavo; Rabossi, Alejandro; Quesada-Allué, Luis A.; Basso, Alicia L.

    2015-01-01

    Abstract The horn fly, Haematobia irritans is an obligate haematophagous cosmopolitan insect pest. The first reports of attacks on livestock by Haematobia irritans in Argentina and Uruguay occurred in 1991, and since 1993 it is considered an economically important pest. Knowledge on the genetic characteristics of the horn fly increases our understanding of the phenotypes resistant to insecticides that repeatedly develop in these insects. The karyotype of Haematobia irritans, as previously described using flies from an inbred colony, shows a chromosome complement of 2n=10 without heterochromosomes (sex chromosomes). In this study, we analyze for the first time the chromosome structure and variation of four wild populations of Haematobia irritans recently established in the Southern Cone of South America, collected in Argentina and Uruguay. In these wild type populations, we confirmed and characterized the previously published “standard” karyotype of 2n=10 without sex chromosomes; however, surprisingly a supernumerary element, called B-chromosome, was found in about half of mitotic preparations. The existence of statistically significant karyotypic diversity was demonstrated through the application of orcein staining, C-banding and H-banding. This study represents the first discovery and characterization of horn fly karyotypes with 2n=11 (2n=10+B). All spermatocytes analyzed showed 5 chromosome bivalents, and therefore, 2n=10 without an extra chromosome. Study of mitotic divisions showed that some chromosomal rearrangements affecting karyotype structure are maintained as polymorphisms, and multiple correspondence analyses demonstrated that genetic variation was not associated with geographic distribution. Because it was never observed during male meiosis, we hypothesize that B-chromosome is preferentially transmitted by females and that it might be related to sex determination. PMID:25893073

  10. Genome-based approach to discover new livestock pest control technologies: The horn fly (Haematobia irritans) genome sequencing project

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Invasion of the Americas by the horn fly, H. irritans, has resulted in control issues throughout the continent. Insecticide resistance is a major complicating factor with efforts to manage this pest. Stakeholder focus groups identified the horn fly as the top priority arthropod pest affecting cattle...

  11. Identification of microorganisms in partially fed female horn flies, Haematobia irritans.

    PubMed

    Torres, Lorena; Almazán, Consuelo; Ayllón, Nieves; Galindo, Ruth C; Rosario-Cruz, Rodrigo; Quiroz-Romero, Héctor; Gortazar, Christian; de la Fuente, José

    2012-09-01

    The horn fly Haematobia irritans (Linnaeus, 1758) (Diptera: Muscidae) is one of the most important ectoparasites of cattle. The parasitism of horn flies interferes with cattle feeding, thus reducing weight gain and milk production. Additionally, horn flies are mechanical vectors of pathogens that cause disease in cattle. The aims of this study were to identify microorganisms in partially fed female horn flies through mining of expressed sequence tags (ESTs) and to characterize microorganism prevalence using real-time RT-PCR. Seven unigenes containing 24 ESTs were homologous to infectious agents. Microorganisms identified in partially fed female horn flies ESTs included Nora virus (3 unigenes; 8 ESTs), Wolbachia endosymbionts (3 unigenes; 3 ESTs), and Mycobacterium bovis (1 unigene; 13 ESTs). These results expanded the repertoire of microorganisms that could cause persistent infections or be mechanically transmitted by horn flies and support further studies on the role of horn flies in the epidemiology of these pathogens in Mexico.

  12. Thrombostasin isoform frequency in a Central Texas field population of the horn fly, Haematobia irritans.

    PubMed

    Untalan, Pia M; Pruett, John H; Atteberry, Heather N; Steelman, C Dayton

    2006-12-20

    Thrombostasin is an anti-thrombin factor that plays a role in successful feeding of the horn fly, Haematobia irritans. It has been isolated and characterized from saliva, and polymorphisms in the gene coding sequence have been previously reported. In the present study, the thrombostasin gene was analyzed from 60, field-collected flies from Camp Stanley, Texas and the allele and genotype frequencies were compared with previously published data for an Alabama field collection and a Texas in vitro colony-reared collection. Significant differences in genotype frequency and extent of genotypic diversity observed between the Alabama and Camp Stanley field collections may be attributable to host genetic differences. In addition, bull calves that were phenotyped as either high- or low-carriers were parasitized by horn flies that displayed a significantly different genotype distribution, supporting a possible explanation for horn fly host selection behavior, as evidenced by thrombostasin sequence analysis of 95 additional horn flies collected from respective hosts.

  13. Influence of environmental and meteorological factors on the biting activity of Leptoconops noei and Leptoconops irritans (Diptera: Ceratopogonidae) in Italy.

    PubMed

    Carrieri, Marco; Montemurro, Eustachio; Valentino, Salvatore Vito; Bellini, Romeo

    2011-03-01

    The flight activity of Leptoconops irritans and L. noei was studied on the Jonian-Lucanian coast of southern Italy, using CO2-baited traps. The flight of the females lasted from 6:00 a.m. to 8:40 p.m., with L. irritans being active in the morning hours while L. noei peaked around 6:00 p.m. Based on a stepwise regression analysis, temperature, RH, solar radiation, trap proximity to larval habitats, and the time of the day seemed to have little influence on the biting cycle of the 2 biting midges. Only a shift in wind direction appeared to influence female dispersion, resulting into population fluctuations of both species.

  14. Dynamics of cypermethrin resistance in the field in the horn fly, Haematobia irritans.

    PubMed

    Guglielmone, A A; Castelli, M E; Volpogni, M M; Anziani, O S; Mangold, A J

    2002-09-01

    The toxicity of cypermethrin to the horn fly Haematobia irritans (L.) (Diptera: Muscidae) was determined for samples collected from untreated herds at a farm in central Argentina from October 1997 to May 2001. Field tests of the efficacy of cypermethrin against horn flies were first carried out at this farm in 1993, when the fly was shown to be susceptible to pyrethroids. Subsequently the horn fly populations on this farm were shown to have become resistant and, since 1997, the use of cypermethrin has been restricted to experimental purposes. In this study, fly samples collected in 1999, 2000 and 2001 were subjected to a polymerase chain reaction (PCR) to detect the presence of a specific nucleotide substitution in the sodium channel gene sequence, which has been associated with target site insensitivity to pyrethroids. This analysis showed that the level of cypermethrin resistance had diminished between 1997 and 2001. However, this was not sufficient to restore the efficacy of this pyrethroid to the level found prior to the onset of resistance. Heterozygous and homozygous resistant flies were detected in all samples of flies subjected to PCR diagnosis of alleles conferring target site resistance.

  15. Pyrosequencing-based analysis of the microbiome associated with the horn fly, Haematobia irritans.

    PubMed

    Palavesam, Azhahianambi; Guerrero, Felix D; Heekin, Andrew M; Wang, Ju; Dowd, Scot E; Sun, Yan; Foil, Lane D; Pérez de León, Adalberto A

    2012-01-01

    The horn fly, Haematobia irritans, is one of the most economically important pests of cattle. Insecticides have been a major element of horn fly management programs. Growing concerns with insecticide resistance, insecticide residues on farm products, and non-availability of new generation insecticides, are serious issues for the livestock industry. Alternative horn fly control methods offer the promise to decrease the use of insecticides and reduce the amount of insecticide residues on livestock products and give an impetus to the organic livestock farming segment. The horn fly, an obligatory blood feeder, requires the help of microflora to supply additional nutrients and metabolize the blood meal. Recent advancements in DNA sequencing methodologies enable researchers to examine the microflora diversity independent of culture methods. We used the bacterial 16S tag-encoded FLX-titanium amplicon pyrosequencing (bTEFAP) method to carry out the classification analysis of bacterial flora in adult female and male horn flies and horn fly eggs. The bTEFAP method identified 16S rDNA sequences in our samples which allowed the identification of various prokaryotic taxa associated with the life stage examined. This is the first comprehensive report of bacterial flora associated with the horn fly using a culture-independent method. Several rumen, environmental, symbiotic and pathogenic bacteria associated with the horn fly were identified and quantified. This is the first report of the presence of Wolbachia in horn flies of USA origin and is the first report of the presence of Rikenella in an obligatory blood feeding insect.

  16. Pyrosequencing-Based Analysis of the Microbiome Associated with the Horn Fly, Haematobia irritans

    PubMed Central

    Palavesam, Azhahianambi; Guerrero, Felix D.; Heekin, Andrew M.; Wang, Ju; Dowd, Scot E.; Sun, Yan; Foil, Lane D.; Pérez de León, Adalberto A.

    2012-01-01

    The horn fly, Haematobia irritans, is one of the most economically important pests of cattle. Insecticides have been a major element of horn fly management programs. Growing concerns with insecticide resistance, insecticide residues on farm products, and non-availability of new generation insecticides, are serious issues for the livestock industry. Alternative horn fly control methods offer the promise to decrease the use of insecticides and reduce the amount of insecticide residues on livestock products and give an impetus to the organic livestock farming segment. The horn fly, an obligatory blood feeder, requires the help of microflora to supply additional nutrients and metabolize the blood meal. Recent advancements in DNA sequencing methodologies enable researchers to examine the microflora diversity independent of culture methods. We used the bacterial 16S tag-encoded FLX-titanium amplicon pyrosequencing (bTEFAP) method to carry out the classification analysis of bacterial flora in adult female and male horn flies and horn fly eggs. The bTEFAP method identified 16S rDNA sequences in our samples which allowed the identification of various prokaryotic taxa associated with the life stage examined. This is the first comprehensive report of bacterial flora associated with the horn fly using a culture-independent method. Several rumen, environmental, symbiotic and pathogenic bacteria associated with the horn fly were identified and quantified. This is the first report of the presence of Wolbachia in horn flies of USA origin and is the first report of the presence of Rikenella in an obligatory blood feeding insect. PMID:23028533

  17. Efficacy of entomopathogenic fungi (Ascomycetes: Hypocreales) against adult Haematobia irritans (Diptera: Muscidae) under stable conditions in the Mexican dry tropics.

    PubMed

    Galindo-Velasco, E; Lezama-Gutiérrez, R; Cruz-Vázquez, C; Pescador-Rubio, A; Angel-Sahagún, C A; Ojeda-Chi, M M; Rodríguez-Vivas, R I; Contreras-Lara, D

    2015-04-30

    The purpose of this paper is to evaluate the effect of five strains of Metarhizium anisopliae (Ma) and three strains of Isaria fumosorosea (Ifr) at a concentration of 1×10(8)colony-forming units/ml applied by spraying onto bovines with controlled infestation of Haematobia irritans under stable conditions in the Mexican dry tropics. Four experiments were performed, in each of which three treatments (two fungal strains and one control) were evaluated with eight repetitions for each one, by carrying out a single application of the aqueous suspension of each strain. The animals were isolated in individual cages and direct counts of the infestation were carried out for 13 days. It was observed that strains Ma2, Ma6, Ma10, Ma14, and Ma34 caused 94-100% reduction in infestation between days 12 and 13 post-treatment, while strains Ifr19, Ifr11, and Ifr12 reduced infestation from 90% to 98% up to day 13 post-application. There was an effect in the generation of horn flies from the excrement of bovines that were treated with different strains, reducing the reproduction of subsequent generations. It was concluded that the strains of M. anisopliae and I. fumosorosea evaluated in this study can be used as biocontrol agents in infestations of H. irritans in stabled bovines.

  18. The effects of hair density of beef cattle on Haematobia irritans horn fly populations.

    PubMed

    Steelman, C D; Brown, M A; Gbur, E E; Tolley, G

    1997-07-01

    We show the relationships that exist between the amount of hair and quantity of sebum on cattle skin and the population density of the horn fly, Haematobia irritans. Brahman and Chianina steers had means of 2390 and 1587 hairs per cm2, respectively, significantly more than the mean number of hairs on Angus, Brahman x Angus Crossbred, Charolais, and Red Poll steers. The Chianina steers had > 30% more sebum present on their skin and hair (0.58g/929 cm2) than the Angus, Charolais, and Red Poll steers at the Beef Cattle Research Station Savoy, Arkansas. The Brahman steers had a significantly greater amount of sebum present on the skin (1.51 g/929 cm2) than the Crossbred and purebred Angus steers (0.55 and 0.25 g/929 cm2, respectively) at the South Central Family Farms Research Centre Booneville, Arkansas. The Brahman and Chianina steers had means of 61.9 and 17.0 horn flies per steer, respectively, during the fly season, whereas the Angus, Crossbred, Charolais and Red Poll steers had fly season means that ranged from 76.9 to 265.8 flies per steer. Regression analysis showed that an increase of 100 hairs per cm2, was associated with a reduction of 11 horn flies in the Angus II, 5 in Angus I, 20 in Charolais, 37 in Red Poll, and 0.4 in Chianina steers at the Savoy Station and a reduction of 6.6 horn flies for the Angus, Brahman, and Crossbred steers at the Booneville Centre. Regardless of cattle breed, an increase of 1.0 g of sebum per 929 cm2 output by the steer was associated with 478.5 additional hairs per cm2 on the animal. Each increase of 0.25 g of sebum per 929 cm2 resulted in a decrease of 9.2 horn flies per steer. We conclude that some of the factors responsible for fly-resistance in cattle are hair density and the corresponding amount of sebum present on cattle skin and hair. PMID:9330257

  19. The effects of hair density of beef cattle on Haematobia irritans horn fly populations.

    PubMed

    Steelman, C D; Brown, M A; Gbur, E E; Tolley, G

    1997-07-01

    We show the relationships that exist between the amount of hair and quantity of sebum on cattle skin and the population density of the horn fly, Haematobia irritans. Brahman and Chianina steers had means of 2390 and 1587 hairs per cm2, respectively, significantly more than the mean number of hairs on Angus, Brahman x Angus Crossbred, Charolais, and Red Poll steers. The Chianina steers had > 30% more sebum present on their skin and hair (0.58g/929 cm2) than the Angus, Charolais, and Red Poll steers at the Beef Cattle Research Station Savoy, Arkansas. The Brahman steers had a significantly greater amount of sebum present on the skin (1.51 g/929 cm2) than the Crossbred and purebred Angus steers (0.55 and 0.25 g/929 cm2, respectively) at the South Central Family Farms Research Centre Booneville, Arkansas. The Brahman and Chianina steers had means of 61.9 and 17.0 horn flies per steer, respectively, during the fly season, whereas the Angus, Crossbred, Charolais and Red Poll steers had fly season means that ranged from 76.9 to 265.8 flies per steer. Regression analysis showed that an increase of 100 hairs per cm2, was associated with a reduction of 11 horn flies in the Angus II, 5 in Angus I, 20 in Charolais, 37 in Red Poll, and 0.4 in Chianina steers at the Savoy Station and a reduction of 6.6 horn flies for the Angus, Brahman, and Crossbred steers at the Booneville Centre. Regardless of cattle breed, an increase of 1.0 g of sebum per 929 cm2 output by the steer was associated with 478.5 additional hairs per cm2 on the animal. Each increase of 0.25 g of sebum per 929 cm2 resulted in a decrease of 9.2 horn flies per steer. We conclude that some of the factors responsible for fly-resistance in cattle are hair density and the corresponding amount of sebum present on cattle skin and hair.

  20. Chemical composition and fumigant toxicity of the essential oils from 16 species of Eucalyptus against Haematobia irritans (Diptera: Muscidae) adults.

    PubMed

    Juan, Laura W; Lucia, Alejandro; Zerba, Eduardo N; Harrand, Leonel; Marco, Martin; Masuh, Hector M

    2011-06-01

    Oils extracted from various species of Eucalyptus (Eucalyptus badjensis Beuzev & Welch, Eucalyptus badjensis x Eucalyptus nitens, Eucalyptus benthamii variety dorrigoensis Maiden & Cambage, Eucalyptus botryoides Smith, Eucalyptus dalrympleana Maiden, Eucalyptus fastigata Deane & Maiden, Eucalyptus nobilis L.A.S. Johnson & K. D. Hill, Eucalyptus polybractea R. Baker, Eucalyptus radiata ssp. radiata Sieber ex Spreng, Eucalyptus resinifera Smith, Eucalyptus robertsonii Blakely, Eucalyptus rubida Deane & Maiden, Eucalyptus smithii R. Baker, Eucalyptus elata Dehnh, Eucalyptus fraxinoides Deane & Maiden, E. obliqua L'Hér) were obtained by hydrodistillation. The chemical composition of essential oils was determined by gas chromatography coupled to mass spectrometry. Essential oils were mainly composed of 1,8-cineole, alpha-pinene, alpha-terpineol, 4-terpineol, and p-cymene. Vapors from these essential oils and their major components were found to be toxic to Haematobia irritans (L.) (Diptera: Muscidae) adults. An aliquot of each oil was placed in a cylindrical test chamber, and the number of knocked down flies was recorded as a function of time. Knockdown time 50% was then calculated. Results showed that essential oil of E. polybractea had the highest knockdown activity of 3.44 min. A correlation was observed between the content of 1,8-cineole in the Eucalyptus essential oils and the corresponding toxic effect.

  1. Chemical composition and fumigant toxicity of the essential oils from 16 species of Eucalyptus against Haematobia irritans (Diptera: Muscidae) adults.

    PubMed

    Juan, Laura W; Lucia, Alejandro; Zerba, Eduardo N; Harrand, Leonel; Marco, Martin; Masuh, Hector M

    2011-06-01

    Oils extracted from various species of Eucalyptus (Eucalyptus badjensis Beuzev & Welch, Eucalyptus badjensis x Eucalyptus nitens, Eucalyptus benthamii variety dorrigoensis Maiden & Cambage, Eucalyptus botryoides Smith, Eucalyptus dalrympleana Maiden, Eucalyptus fastigata Deane & Maiden, Eucalyptus nobilis L.A.S. Johnson & K. D. Hill, Eucalyptus polybractea R. Baker, Eucalyptus radiata ssp. radiata Sieber ex Spreng, Eucalyptus resinifera Smith, Eucalyptus robertsonii Blakely, Eucalyptus rubida Deane & Maiden, Eucalyptus smithii R. Baker, Eucalyptus elata Dehnh, Eucalyptus fraxinoides Deane & Maiden, E. obliqua L'Hér) were obtained by hydrodistillation. The chemical composition of essential oils was determined by gas chromatography coupled to mass spectrometry. Essential oils were mainly composed of 1,8-cineole, alpha-pinene, alpha-terpineol, 4-terpineol, and p-cymene. Vapors from these essential oils and their major components were found to be toxic to Haematobia irritans (L.) (Diptera: Muscidae) adults. An aliquot of each oil was placed in a cylindrical test chamber, and the number of knocked down flies was recorded as a function of time. Knockdown time 50% was then calculated. Results showed that essential oil of E. polybractea had the highest knockdown activity of 3.44 min. A correlation was observed between the content of 1,8-cineole in the Eucalyptus essential oils and the corresponding toxic effect. PMID:21735933

  2. The discovery of isoxazoline oxime ethers as a new class of ectoparasiticides for the control of Haematobia irritans (horn fly) in cattle.

    PubMed

    Curtis, Michael P; Chubb, Nathan; Ellsworth, Edmund; Goodwin, Richard; Holzmer, Sue; Koch, Jason; McTier, Tom; Menon, Sanjay; Mills, Kent; Pullins, Aleah; Stuk, Tim; Zinser, Erich

    2014-11-01

    Haematobia irritans (horn fly) infestation in cattle is responsible for over a billion dollars a year in global economic loss due to decreased milk production and lower feed conversion. There is significant need for new insecticidal agents since current treatments such as organophosphates and pyrethroids suffer from field resistance. Isoxazoline oxime ethers represent a new class of γ-aminobutyric acid (GABA) receptor channel blockers which show good activity (LD(90) = 1.0 μg/mL) against horn flies in an in vitro feed assay and have demonstrated efficacy (>90% reduction at 1.0mg/kg) as a topical treatment in a field study.

  3. Variation in the load of the horn fly, Haematobia irritans, in cattle herds is determined by the presence or absence of individual heifers.

    PubMed

    Jensen, K-M V; Jespersen, J B; Birkett, M A; Pickett, J A; Thomas, G; Wadhams, L J; Woodcock, C M

    2004-09-01

    The distribution of horn flies, Haematobia irritans (L.) (Diptera: Muscidae), in herds of Danish Holstein-Friesian cattle was investigated in two studies conducted during two field seasons. In the first study, highly significant differences in fly distribution between the most and the least fly-susceptible heifers were observed. In one herd, the mean difference between the most fly-susceptible and the most fly-resistant heifers was 268 Ha. irritans specimens. The highest ratio between upper and lower mean fly number was 64.1:1, whereas the lowest was 3.1:1. In the second year, it was demonstrated that the heifers kept their rank in fly attraction over time. The trial clearly demonstrated that some heifers were attracting flies, whereas others, even in the same herd, only carried a few. In the second study, heifers were moved in and out of herds in an attempt to manipulate fly loads in the herds. In year 1, one herd (herd A) received four fly-resistant heifers from another herd (herd B), resulting in a drop in the mean number of flies, whereas herd B received four fly-susceptible heifers from herd A, resulting in an elevation of the mean number of flies. In year 2, a similar pattern emerged using herds C and D, and when the cattle were later returned to their original herds, the fly loads returned to their original distribution. The data presented here show unequivocally that, for horn flies, there can be considerable differences in fly loads for individual heifers within the Holstein-Friesian breed. Furthermore, the overall fly load within herds can be manipulated, and can be reversed. Thus, the distribution in the number of flies within a herd appears to depend on the number of fly-resistant or fly-susceptible heifers. The possible role of chemical factors emitted by heifers, i.e. volatile semiochemicals, in determining differences in fly loads is discussed, whereby attractants are emitted by fly-susceptible heifers and enable flies to locate their host, and

  4. Effect of diflubenzuron on immature stages of Haematobia irritans (L.) (Diptera: Muscidae) in Uberlândia, State of Minas Gerais, Brazil.

    PubMed

    Silva, Juliana Junqueira da; Mendes, Júlio

    2002-07-01

    Horn fly immatures were raised in media containing different concentrations of diflubenzuron in order to verify their susceptibility to this insect growth regulator (IGR). The 50% and 95% lethal concentrations of diflubenzuron for the population (LC50, LC95) were determined as well as the effect of this IGR on the different immature horn fly stages. The tests were performed using the progeny of adults collected in the field. The immatures were maintained in a growth chamber at 25.0 +/- 0.5oC and 12-12 h photoperiod. IGR concentrations of 300 ppb, 100 ppb and 50 ppb were lethal for 100% of the sample. Pupae malformation occurred in the breeding media containing different diflubenzuron concentrations. Values for LC50, LC95 (+/- 95% fiducial limits) and the slope of the regression line were respectively, 25.521 +/- 1.981 ppb, 34.650 +/- 2.001 ppb and 12.720 +/- 1.096. The third larval instar was more sensitive to the sub-lethal concentration of the product than the first and second ones were. The results indicate that this IGR can be an important tool for controlling horn fly populations as well as for managing horn fly resistance to conventional insecticides against Haematobia irritans in Uberlândia, State of Minas Gerais.

  5. Molecular characterization and expression analysis of interferon-gamma in the large yellow croaker Larimichthys crocea.

    PubMed

    Chen, Ruan-Ni; Su, Yong-Quan; Wang, Jun; Liu, Min; Qiao, Ying; Mao, Yong; Ke, Qiao-Zhen; Han, Kun-Huang; Zheng, Wei-Qiang; Zhang, Jian-She; Wu, Chang-Wen

    2015-10-01

    The large yellow croaker Larimichthys crocea is an important mariculture fish species in China, and the bacterium Vibrio harveyi (V. harveyi) and the ciliate protozoan Cryptocaryon irritans (C. irritans) are the two major pathogens in its aquaculture sector. Interferon-gamma (IFN-γ) plays important roles in regulating both innate and cell mediated immune responses as an inflammatory cytokine. In this study, we obtained the nucleotide sequence of IFN-γ from the large yellow croaker (LcIFN-γ). The phylogenetic relationship tree of 18 available IFN-γ genes was constructed based on their sequences. Expression analyses in 10 various tissues were conducted after the croaker challenged with V. harveyi and C. irritans, respectively. Real time PCR analysis showed that the expression of LcIFN-γ was observed broadly in health individuals. After injected with V. harveyi, the 10 tissues had a higher expression of IFN-γ at the first day (1 d); only spleen, muscle, intestine, heart and skin had higher expressions after infected with C. irritans at 1 d. Major immune tissues (skin, gill, head kidney and spleen) and detoxification tissue (liver) were sampled at 0 h, 6 h, 1 d, 2 d, 3 d, 4 d, 5 d and 7 d to understand the expression trends of LcIFN-γ after challenged with C. irritans. The expressions of LcIFN-γ in skin and gill (the primary immune organs) showed a clear correlative relationship with the life cycle of C. irritans.

  6. Susceptibility of the horn fly, Haematobia irritans irritans (Diptera: Muscidae), to insecticides in Brazil.

    PubMed

    Barros, Antonio Thadeu Medeiros; Saueressig, Thelma Maria; Gomes, Alberto; Koller, Wilson Werner; Furlong, John; Girão, Eneide Santiago; Pinheiro, Alfredo da Cunha; Alves-Branco, Francisco de Paula Jardim; Sapper, Maria de Fátima Munhós; Braga, Ramayana Menezes; Oliveira, Amaury Apolonio de

    2012-01-01

    Since horn fly populations became established throughout Brazil, complaints regarding control failure have increased around the country. A broad survey to evaluate the susceptibility of horn flies to both organophosphate (OP) and pyrethroid insecticides was conducted from October 2000 to April 2003. Bioassays using filter papers impregnated with cypermethrin, permethrin or diazinon were conducted on 154 horn fly populations in 14 states and 78 municipalities. Resistance to cypermethrin, the active ingredient present in most insecticide products for horn fly control in Brazil, was detected in 98.46% of the populations, with resistance ratios (RR) ranging from 2.5 to 719.9. Resistance to permethrin (RRs < 6.3) was found in 96.67% of the populations, despite its lack of use. In general, pyrethroid resistance was detected in 97.18% of the horn fly populations, with frequencies greater than 87% in all regions of the country. The status of susceptibility of horn fly populations in Brazil to insecticides can be characterized by high susceptibility to OPs and widespread resistance to pyrethroids, potentially compromising the efficacy of pyrethroid products in most cases. Although some partial results have previously been presented, a general picture of horn fly susceptibility in Brazil is presented here for the first time.

  7. Susceptibility of the horn fly, Haematobia irritans irritans (Diptera: Muscidae), to insecticides in Brazil.

    PubMed

    Barros, Antonio Thadeu Medeiros; Saueressig, Thelma Maria; Gomes, Alberto; Koller, Wilson Werner; Furlong, John; Girão, Eneide Santiago; Pinheiro, Alfredo da Cunha; Alves-Branco, Francisco de Paula Jardim; Sapper, Maria de Fátima Munhós; Braga, Ramayana Menezes; Oliveira, Amaury Apolonio de

    2012-01-01

    Since horn fly populations became established throughout Brazil, complaints regarding control failure have increased around the country. A broad survey to evaluate the susceptibility of horn flies to both organophosphate (OP) and pyrethroid insecticides was conducted from October 2000 to April 2003. Bioassays using filter papers impregnated with cypermethrin, permethrin or diazinon were conducted on 154 horn fly populations in 14 states and 78 municipalities. Resistance to cypermethrin, the active ingredient present in most insecticide products for horn fly control in Brazil, was detected in 98.46% of the populations, with resistance ratios (RR) ranging from 2.5 to 719.9. Resistance to permethrin (RRs < 6.3) was found in 96.67% of the populations, despite its lack of use. In general, pyrethroid resistance was detected in 97.18% of the horn fly populations, with frequencies greater than 87% in all regions of the country. The status of susceptibility of horn fly populations in Brazil to insecticides can be characterized by high susceptibility to OPs and widespread resistance to pyrethroids, potentially compromising the efficacy of pyrethroid products in most cases. Although some partial results have previously been presented, a general picture of horn fly susceptibility in Brazil is presented here for the first time. PMID:22832752

  8. Soluble interleukin-1 receptor, a potential negative regulator of orange-spotted grouper Epinephelus coioides interleukin-1 system.

    PubMed

    Lu, D Q; Yao, M; Yi, S B; Li, Y W; Liu, X C; Zhang, Y; Lin, H R

    2013-09-01

    In this study, the cDNA sequence encoding interleukin-1 (Il-1) receptor-like protein of orange-spotted grouper Epinephelus coioides was obtained. The newly identified sequence was named soluble type I Il-1 receptor (sIl-1rI) owing to its structural composition, which had two Ig-like domains, lack of transmembrane region and the Toll/interleukin-1 receptor (TIR) domain, similar to the brown rat Rattus norvegicus soluble Il-1rI. In addition, sequence comparison and phylogenetic analysis indicated that E. coioides sequence had a closer relationship with Il-1rI than Il-1rII. Real-time PCR revealed that sil-1rI mRNA expression presented a process of decrease, restoration and increase in Cryptocaryon irritans-infected E. coioides. The negative correlation between Il-1β and sil-1rI mRNA in C. irritans-infected head-kidney implied the potential negative regulatory role of sil-1rI in E. coioides Il-1 system. The leucocytes incubated with lipopolysaccharide or polyriboinosinic polyribocytidylic acid exhibited different expression profiles of sil-1rI. Recombinant Il-1β (rIl-1β) protein was capable of inducing sil-1rI mRNA under the concentration of 100 ng ml(-1) , suggesting that high dosage or excess Il-1β would stimulate the expression of sil-1rI to maintain the homoeostasis of E. coioides Il-1 system. For the first time, the role of teleost Il-1rI in parasite infection has been identified, and soluble Il-1r was found in fish.

  9. Ciliate diversity, community structure, and novel taxa in lakes of the McMurdo Dry Valleys, Antarctica.

    PubMed

    Xu, Yuan; Vick-Majors, Trista; Morgan-Kiss, Rachael; Priscu, John C; Amaral-Zettler, Linda

    2014-10-01

    We report an in-depth survey of next-generation DNA sequencing of ciliate diversity and community structure in two permanently ice-covered McMurdo Dry Valley lakes during the austral summer and autumn (November 2007 and March 2008). We tested hypotheses on the relationship between species richness and environmental conditions including environmental extremes, nutrient status, and day length. On the basis of the unique environment that exists in these high-latitude lakes, we expected that novel taxa would be present. Alpha diversity analyses showed that extreme conditions-that is, high salinity, low oxygen, and extreme changes in day length-did not impact ciliate richness; however, ciliate richness was 30% higher in samples with higher dissolved organic matter. Beta diversity analyses revealed that ciliate communities clustered by dissolved oxygen, depth, and salinity, but not by season (i.e., day length). The permutational analysis of variance test indicated that depth, dissolved oxygen, and salinity had significant influences on the ciliate community for the abundance matrices of resampled data, while lake and season were not significant. This result suggests that the vertical trends in dissolved oxygen concentration and salinity may play a critical role in structuring ciliate communities. A PCR-based strategy capitalizing on divergent eukaryotic V9 hypervariable region ribosomal RNA gene targets unveiled two new genera in these lakes. A novel taxon belonging to an unknown class most closely related to Cryptocaryon irritans was also inferred from separate gene phylogenies.

  10. Ciliate diversity, community structure, and novel taxa in lakes of the McMurdo Dry Valleys, Antarctica.

    PubMed

    Xu, Yuan; Vick-Majors, Trista; Morgan-Kiss, Rachael; Priscu, John C; Amaral-Zettler, Linda

    2014-10-01

    We report an in-depth survey of next-generation DNA sequencing of ciliate diversity and community structure in two permanently ice-covered McMurdo Dry Valley lakes during the austral summer and autumn (November 2007 and March 2008). We tested hypotheses on the relationship between species richness and environmental conditions including environmental extremes, nutrient status, and day length. On the basis of the unique environment that exists in these high-latitude lakes, we expected that novel taxa would be present. Alpha diversity analyses showed that extreme conditions-that is, high salinity, low oxygen, and extreme changes in day length-did not impact ciliate richness; however, ciliate richness was 30% higher in samples with higher dissolved organic matter. Beta diversity analyses revealed that ciliate communities clustered by dissolved oxygen, depth, and salinity, but not by season (i.e., day length). The permutational analysis of variance test indicated that depth, dissolved oxygen, and salinity had significant influences on the ciliate community for the abundance matrices of resampled data, while lake and season were not significant. This result suggests that the vertical trends in dissolved oxygen concentration and salinity may play a critical role in structuring ciliate communities. A PCR-based strategy capitalizing on divergent eukaryotic V9 hypervariable region ribosomal RNA gene targets unveiled two new genera in these lakes. A novel taxon belonging to an unknown class most closely related to Cryptocaryon irritans was also inferred from separate gene phylogenies. PMID:25411375

  11. Impact and control of protozoan parasites in maricultured fishes.

    PubMed

    Buchmann, Kurt

    2015-01-01

    Aquaculture, including both freshwater and marine production, has on a world scale exhibited one of the highest growth rates within animal protein production during recent decades and is expected to expand further at the same rate within the next 10 years. Control of diseases is one of the most prominent challenges if this production goal is to be reached. Apart from viral, bacterial, fungal and metazoan infections it has been documented that protozoan parasites affect health and welfare and thereby production of fish in marine aquaculture. Representatives within the main protozoan groups such as amoebae, dinoflagellates, kinetoplastid flagellates, diplomonadid flagellates, apicomplexans, microsporidians and ciliates have been shown to cause severe morbidity and mortality among farmed fish. Well studied examples are Neoparamoeba perurans, Amyloodinium ocellatum, Spironucleus salmonicida, Ichthyobodo necator, Cryptobia salmositica, Loma salmonae, Cryptocaryon irritans, Miamiensis avidus and Trichodina jadranica. The present report provides details on the parasites' biology and impact on productivity and evaluates tools for diagnosis, control and management. Special emphasis is placed on antiprotozoan immune responses in fish and a strategy for development of vaccines is presented.

  12. Optimal protein extraction methods from diverse sample types for protein profiling by using Two-Dimensional Electrophoresis (2DE).

    PubMed

    Tan, A A; Azman, S N; Abdul Rani, N R; Kua, B C; Sasidharan, S; Kiew, L V; Othman, N; Noordin, R; Chen, Y

    2011-12-01

    There is a great diversity of protein samples types and origins, therefore the optimal procedure for each sample type must be determined empirically. In order to obtain a reproducible and complete sample presentation which view as many proteins as possible on the desired 2DE gel, it is critical to perform additional sample preparation steps to improve the quality of the final results, yet without selectively losing the proteins. To address this, we developed a general method that is suitable for diverse sample types based on phenolchloroform extraction method (represented by TRI reagent). This method was found to yield good results when used to analyze human breast cancer cell line (MCF-7), Vibrio cholerae, Cryptocaryon irritans cyst and liver abscess fat tissue. These types represent cell line, bacteria, parasite cyst and pus respectively. For each type of samples, several attempts were made to methodically compare protein isolation methods using TRI-reagent Kit, EasyBlue Kit, PRO-PREP™ Protein Extraction Solution and lysis buffer. The most useful protocol allows the extraction and separation of a wide diversity of protein samples that is reproducible among repeated experiments. Our results demonstrated that the modified TRI-reagent Kit had the highest protein yield as well as the greatest number of total proteins spots count for all type of samples. Distinctive differences in spot patterns were also observed in the 2DE gel of different extraction methods used for each type of sample. PMID:22433892

  13. Characterization and expression analysis of BCA2 gene in large yellow croaker, Larimichthys crocea.

    PubMed

    Zhang, Dong Ling; Yu, Da Hui; Chen, Jian; Wang, Zhi Yong

    2015-10-01

    BCA2, as an E3 ubiquitin ligase, is an important anti-virus immune factor in mammals. Up to date, there are not any related reports on BCA2 protein in fishes yet. In the present investigation, BCA2 in large yellow croaker Larimichthys crocea (named as LcBCA2) was identified and characterized. The full-length cDNA of LcBCA2 was 1571 bp, including an ORF of 888 bp encoding a polypeptide of 295 amino acids. The putative LcBCA2 protein contained a RING-H2 motif at C terminal. The LcBCA2 transcripts were broadly distributed in all detected tissues, with high expression in muscle, moderate in blood, skin, heart, liver and spleen, weak in other tissue as indicated by qPCR analysis. Significant increases were observed in skin, gill and spleen after infection of Cryptocaryon irritans, and in spleen and head-kidney after inactivated Vibrio. parahaemolyticus, LPS and Poly I:C stimulations. Tissue localization by in-situ hybridization showed that LcBCA2 mainly expressed in the spleen of the fish in the test group. Our findings showed that LcBCA2 inclined to sharply increase in immune organs, especially in head-kidney after bacterial and viral stimulations, while in locations (skin and gill) of parasites infections, suggesting that BCA2 may play an important role in fish defense against bacteria, virus and parasites infections, but the immune mechanisms is are different.

  14. The serum of rabbitfish (Siganus oramin) has antimicrobial activity to some pathogenic organisms and a novel serum L-amino acid oxidase is isolated.

    PubMed

    Wang, Fanghua; Li, Ruijun; Xie, Mingquan; Li, Anxing

    2011-01-01

    The serum of rabbitfish (Siganus oramin) has been confirmed previously to have killing effect to Cryptocaryon irritans, an important marine ciliate protozoan that causes a disease referred to as "marine white spot disease". Herein, we find the serum of the rabbitfish also shows antibacterial activity against both gram-positive and gram-negative bacteria and has killing effect on two other parasites: Trypanosoma brucei brucei, Ichthyophthirius multifiliis. Results of scanning electron microscopy indicated that after treating with rabbitfish serum, the surface of the Staphylococcus aureus was wrinkled and pores were formed on the surface of Escherichia coli. Serum of the rabbitfish possesses a strong killing effect to Ichthyophthirius multifiliis in vitro, causing a similar effect as to C. irritans. The serum of rabbitfish also showed strong killing effect to T. b. brucei in vitro, with the minimus trypanocidal titre (MTT) only to be 1.5% in 1 h. Results of laser confocal fluorescence microscopy indicated that rabbitfish serum could also induce cell rupture of T. b. brucei. A novel antimicrobial protein (SR-LAAO) was isolated from the serum of rabbitfish by using ultrafiltration, reversed phase high performance liquid chromatography (RP-HPLC) and Native polyacrylamide gel electrophoresis (Native-PAGE). Results of gel overlay assay showed that the protein could act alone to inhibit the growth of S. aureus and E. coli. Results of western blot and automated Edman degradation showed that it was the same as the antiparasitic protein (APP) reported before to have killing effect on C. irritans. Full length cDNA sequence of the SR-LAAO was cloned. BLAST research suggested that the cDNA of SR-LAAO has a close similarity with a number of L-amino acid oxidases (LAAOs) and possesses two conserved motifs that exist in LAAOs. Combined, these results demonstrate that this protein which has antimicrobial activity to some pathogenic organisms was a novel LAAO found in the serum of

  15. Immobilization antigen vaccine adjuvanted by parasitic heat shock protein 70C confers high protection in fish against cryptocaryonosis.

    PubMed

    Josepriya, T A; Chien, Kuo-Hsuan; Lin, Hsin-Yun; Huang, Han-Ning; Wu, Chang-Jer; Song, Yen-Ling

    2015-08-01

    The immobilization antigen (iAg) has been demonstrated as a protective immunogen against Cryptocaryon irritans infection. In this study, C-terminal domain of heat shock protein 70 cloned from C. irritans (Hsp70C) was tested for its immuno-stimulatory effects. The iAg and Hsp70C cDNAs were constructed independently in secretory forms and were encapsulated in chitosan nanoparticles. In the first immunization trial, grouper fingerlings orally intubated with iAg and iAg:Hsp70C presented 96% and 100% relative percent survival (RPS), respectively, after a lethal challenge. In the second trial, both iAg and iAg:Hsp70C groups showed 100% RPS and the skin trophont burden was significantly lowered. The iAg:Hsp70C still provides a significantly high protection of 51% RPS at 49 days post immunization, when an even more serious lethal infection occurs. RT-qPCR results showed that Hsp70C could up-regulate the expression of i) T cell markers: Cluster of Differentiation 8 alpha (CD8α) and CD4, ii) cytokine genes: Interferon gamma (IFNγ), Tumor Necrosis Factor alpha (TNFα) and Interleukin 12 p40 (IL-12/P40), iii) antibody genes: Immunoglobulin M heavy chain (IgMH) and IgTH, and iv) major histocompatibility complex (MHC-I & MHC-II), in the spleen of iAg:Hsp70C group. Furthermore, significantly high levels of iAg-specific IgM was detected in skin mucus which efficiently immobilized live theronts in iAg- and iAg:Hsp70C-immunized fish at 5 weeks post immunization. Hsp70C significantly increased the number of nonspecific CD8(+) skin leucocytes which exerted cytotoxicity against theronts, although cytotoxic activity showed no difference among the various groups. Because of this complementary cooperation of cellular and humoral immune responses, Hsp70C enhances the efficacy of iAg vaccine and constrains C. irritans infection. In view of the severe loss caused by cryptocaryonosis, application of this parasitic vaccine in farmed and ornamental fish, is worthy to be considered. PMID

  16. Molecular characterization and immunolocalization of the olfactory co-recepter Orco from two blood-feeding muscid flies, the stable fly (Stomoxys calcitrans, L.) and the horn fly (Haematobia irritans irritans, L.)

    PubMed Central

    Olafson, Pia Untalan

    2012-01-01

    Biting flies are economically important, blood-feeding pests of medical and veterinary significance. Chemosensory-based biting fly behaviors, such as host/nutrient source localization and ovipositional site selection, are intriguing targets for the development of supplemental control strategies. In an effort to expand our understanding of biting fly chemosensory pathways, transcripts encoding the highly conserved insect odorant co-receptor (Orco) were isolated from two representative biting fly species, the stable fly (Scal\\Orco) and the horn fly (Hirr\\Orco). Orco forms a complex with an odor-specific odorant receptor to form an odor-gated ion channel. The biting fly transcripts were predicted to encode proteins with 87% – 94% amino acid similarity to published insect Orco sequences and were detected in various immature stages as well as in adult structures associated with olfaction, i.e. antennae and maxillary palps, and gustation, i.e. proboscis. Further, the relevant proteins were immunolocalized to specific antennal sensilla using anti-serum raised against a peptide sequence conserved between the two fly species. Results from this study provide a basis for functional evaluation of repellent/attractant effects on as yet uncharacterized stable fly and horn fly conventional odorant receptors. PMID:23278866

  17. Comparisons of antifeedancy and spatial repellency of three natural product repellents against horn flies, Haematobia irritans (Diptera: Muscidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    BACKGROUND: Horn flies are among the most important biting fly pests of cattle in the United States. Horn fly management is largely dependent upon pesticides, which ultimately leads to the rapid development of insecticide resistance. Alternative control strategies, including repellents, have shown p...

  18. Use of electroporation as an option to transform the Horn Fly, Haematobia irritans: a species recalcitrant to microinjection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Horn flies are serious pests of cattle in North America and control of these flies has primarily relied on insecticides. However, the heavy use of insecticides has led to the development of resistance in horn flies and novel methods of fly control are greatly needed. The use of transformation techno...

  19. Identification and characterization of three TLR1 subfamily members from the orange-spotted grouper, Epinephelus coioides.

    PubMed

    Li, Yan-Wei; Xu, Dong-Dong; Li, Xia; Mo, Ze-Quan; Luo, Xiao-Chun; Li, An-Xing; Dan, Xue-Ming

    2016-08-01

    Toll-like receptors (TLRs), which play important roles in host defense against pathogen infection, are the most intensively studied pattern recognition receptors (PRRs). In this study, we identified three novel TLR1 subfamily members, including TLR1 (EcTLR1b), TLR2 (EcTLR2b) and TLR14 (EcTLR14), from the orange-spotted grouper (Epinephelus coioides). EcTLR1b and EcTLR2b displayed low sequence identity with the previously reported grouper TLR1 (EcTLR1a) and TLR2 (EcTLR2a), respectively. The open reading frames (ORFs) of EcTLR1b, EcTLR2b and EcTLR14 contain 2484 bp, 2394 bp and 2640 bp, which encode the corresponding 827 amino acids (aa), 797 aa and 879 aa, respectively. All three TLRs have leucine-rich repeat (LRR) domains (including an LRR-NT (except for EcTLR1b), several LRR motifs and an LRR-CT), a trans-membrane region and a Toll/interleukin-1 receptor (TIR) domain. The TIR domains of the three TLRs exhibited conserved boxes, namely box1, box2 and box3, and their 3D models were similar to those of human TLR1 or TLR2. Sequence alignment demonstrated that the TIR domains of the three TLRs shared higher sequence identity with those of other species than the full-length receptors. Phylogenetic analysis indicated that EcTLR1s and EcTLR2s are characterized by their differing evolutionary status, whereas EcTLR14 was found to be in the same group as other piscine TLR14/18s. The three TLRs were ubiquitously expressed in seven tested tissues of healthy groupers, although their expression profiles were different. Post Cryptocaryon irritans infection, TLR1s expression was up-regulated in the gills. The expression of TLR2b was mainly increased in the spleen, but decreased in the gills, which was similar to the expression pattern of TLR2a post C. irritans infection. Unlike EcTLR1b and EcTLR2b, however, the grouper TLR14 transcript was substantially induced in both tissues post challenge. These findings may be helpful in understanding the innate immune mechanism of host

  20. Identification and characterization of three TLR1 subfamily members from the orange-spotted grouper, Epinephelus coioides.

    PubMed

    Li, Yan-Wei; Xu, Dong-Dong; Li, Xia; Mo, Ze-Quan; Luo, Xiao-Chun; Li, An-Xing; Dan, Xue-Ming

    2016-08-01

    Toll-like receptors (TLRs), which play important roles in host defense against pathogen infection, are the most intensively studied pattern recognition receptors (PRRs). In this study, we identified three novel TLR1 subfamily members, including TLR1 (EcTLR1b), TLR2 (EcTLR2b) and TLR14 (EcTLR14), from the orange-spotted grouper (Epinephelus coioides). EcTLR1b and EcTLR2b displayed low sequence identity with the previously reported grouper TLR1 (EcTLR1a) and TLR2 (EcTLR2a), respectively. The open reading frames (ORFs) of EcTLR1b, EcTLR2b and EcTLR14 contain 2484 bp, 2394 bp and 2640 bp, which encode the corresponding 827 amino acids (aa), 797 aa and 879 aa, respectively. All three TLRs have leucine-rich repeat (LRR) domains (including an LRR-NT (except for EcTLR1b), several LRR motifs and an LRR-CT), a trans-membrane region and a Toll/interleukin-1 receptor (TIR) domain. The TIR domains of the three TLRs exhibited conserved boxes, namely box1, box2 and box3, and their 3D models were similar to those of human TLR1 or TLR2. Sequence alignment demonstrated that the TIR domains of the three TLRs shared higher sequence identity with those of other species than the full-length receptors. Phylogenetic analysis indicated that EcTLR1s and EcTLR2s are characterized by their differing evolutionary status, whereas EcTLR14 was found to be in the same group as other piscine TLR14/18s. The three TLRs were ubiquitously expressed in seven tested tissues of healthy groupers, although their expression profiles were different. Post Cryptocaryon irritans infection, TLR1s expression was up-regulated in the gills. The expression of TLR2b was mainly increased in the spleen, but decreased in the gills, which was similar to the expression pattern of TLR2a post C. irritans infection. Unlike EcTLR1b and EcTLR2b, however, the grouper TLR14 transcript was substantially induced in both tissues post challenge. These findings may be helpful in understanding the innate immune mechanism of host

  1. Expression profiles and interaction suggest TBK1 can be regulated by Nrdp1 in response to immune stimulation in large yellow croaker Larimichthys crocea.

    PubMed

    Zhang, Dong Ling; Yu, Da Hui; Chen, Jian; Fan, Sigang; Wang, Zhi Yong

    2015-10-01

    TBK1 has been extensively studied in mammals because of its important roles as a molecular bridge, linking the TLRs (TLR3 and TLR4) and RLRs signals to activate transcriptional factors IRF3 and IRF7 for IFN-I production. However, the information on molecular and functional characteristics of TBK1 in teleosts is limited. In this study, the molecular characterization and immune response of TBK1 in Larimichthys crocea (named as LcTBK1) as well as its interaction with Nrdp1 were investigated. Sequence analysis demonstrated that LcTBK1 included four functional motifs, the N-terminal protein kinase domain and ATP-binding site, middle ULD and C-terminal coiled-coil domain. The tissue expression profiles indicated that LcTBK1 gene was constitutively expressed in the twelve tissues examined, with high expression in brain. Temporal expression analysis showed that LcTBK1 mRNA was obviously increased in the liver after injection of LPS, Poly I:C and inactive Vibrio parahaemolyticus, however, declined at some time points in spleen and head-kidney. Furthermore, we found that LcTBK1 can interact with LcNrdp1, an E3 ubiquitin ligase that involved in immune response to Cryptocaryon irritans infection in L. crocea. The qPCR showed that LcNrdp1 was also significantly up-regulated in liver, down-regualted at some time points in spleen and head-kidney after LPS, Poly I:C and inactive V. parahaemolyticus injection, although the expression patterns of the two genes after the three treatments were different in change magnitude and up-regulation timespan. These results suggested that LcTBK1 was involved in L. crocea defense against the pathogen infection and can be regulated by Nrdp1 in PPRs signaling pathway of fishes.

  2. Antibacterial efficacy of recombinant Siganus oramin L-amino acid oxidase expressed in Pichia pastoris.

    PubMed

    Li, Ruijun; Li, Anxing

    2014-12-01

    Siganus oraminl-amino acid oxidase is a novel natural protein (named SR-LAAO) isolated from serum of the rabbitfish (S. oramin), which showed antibacterial activity against both Gram-positive and Gram-negative bacteria and had a lethal effect on the parasites Cryptocaryon irritans, Trypanosoma brucei brucei and Ichthyophthirius multifiliis. In order to test whether recombinant SR-LAAO (rSR-LAAO) produced by the eukaryotic expression system also has antimicrobial activity, the yeast Pichia pastoris was used as the expression host to obtain rSR-LAAO in vitro. Crude rSR-LAAO produced by P. pastoris integrated with the SR-LAAO gene had antibacterial activity against both Gram-positive and Gram-negative bacteria as shown by inhibition zone assay of the antibacterial spectrum on agar plates. The average diameter of the inhibition zone of crude rSR-LAAO against the Gram-positive bacteria Staphylococcus aureus and Streptococcus agalactiae was 1.040 ± 0.045 cm and 1.209 ± 0.085 cm, respectively. For the Gram-negative bacteria Aeromonas sobria, Escherichia coli, Vibrio alginolyticus, Vibrio cholera and Photobacterium damselae subsp. piscicida, the average diameter of inhibition zone was 1.291 ± 0.089 cm, 0.943 ± 0.061 cm, 0.756 ± 0.057 cm, 0.834 ± 0.023 cm and 1.211 ± 0.026 cm, respectively. These results were obtained at the logarithmic growth phase of S. agalactiae and A. sobria cell suspensions after incubation with 0.5 mg/mL crude rSR-LAAO for 24 h. The final bacterial growth rate was decreased significantly. The relative inhibition rate can reach 50% compared to crude products from P. pastoris integrated with an empty vector at the same concentration of protein. The antimicrobial activity of crude rSR-LAAO was likely associated with H2O2 formation, because its inhibition zones were disturbed significantly by catalase. Scanning electron microscopy results showed crude rSR-LAAO-treated bacterial surfaces became rough and particles were attached, cell walls were

  3. Antibacterial efficacy of recombinant Siganus oramin L-amino acid oxidase expressed in Pichia pastoris.

    PubMed

    Li, Ruijun; Li, Anxing

    2014-12-01

    Siganus oraminl-amino acid oxidase is a novel natural protein (named SR-LAAO) isolated from serum of the rabbitfish (S. oramin), which showed antibacterial activity against both Gram-positive and Gram-negative bacteria and had a lethal effect on the parasites Cryptocaryon irritans, Trypanosoma brucei brucei and Ichthyophthirius multifiliis. In order to test whether recombinant SR-LAAO (rSR-LAAO) produced by the eukaryotic expression system also has antimicrobial activity, the yeast Pichia pastoris was used as the expression host to obtain rSR-LAAO in vitro. Crude rSR-LAAO produced by P. pastoris integrated with the SR-LAAO gene had antibacterial activity against both Gram-positive and Gram-negative bacteria as shown by inhibition zone assay of the antibacterial spectrum on agar plates. The average diameter of the inhibition zone of crude rSR-LAAO against the Gram-positive bacteria Staphylococcus aureus and Streptococcus agalactiae was 1.040 ± 0.045 cm and 1.209 ± 0.085 cm, respectively. For the Gram-negative bacteria Aeromonas sobria, Escherichia coli, Vibrio alginolyticus, Vibrio cholera and Photobacterium damselae subsp. piscicida, the average diameter of inhibition zone was 1.291 ± 0.089 cm, 0.943 ± 0.061 cm, 0.756 ± 0.057 cm, 0.834 ± 0.023 cm and 1.211 ± 0.026 cm, respectively. These results were obtained at the logarithmic growth phase of S. agalactiae and A. sobria cell suspensions after incubation with 0.5 mg/mL crude rSR-LAAO for 24 h. The final bacterial growth rate was decreased significantly. The relative inhibition rate can reach 50% compared to crude products from P. pastoris integrated with an empty vector at the same concentration of protein. The antimicrobial activity of crude rSR-LAAO was likely associated with H2O2 formation, because its inhibition zones were disturbed significantly by catalase. Scanning electron microscopy results showed crude rSR-LAAO-treated bacterial surfaces became rough and particles were attached, cell walls were

  4. Molecular characterization and functional analysis of TRAF6 in orange-spotted grouper (Epinephelus coioides).

    PubMed

    Li, Yan-Wei; Li, Xia; Xiao, Xi-Xi; Zhao, Fei; Luo, Xiao-Chun; Dan, Xue-Ming; Li, An-Xing

    2014-05-01

    Tumor necrosis factor receptor (TNFR)-associated factor 6 (TRAF6) is a crucial signal transducer in both the TNFR superfamily and Toll-like receptor/interleukin 1R family. Although significant progress has been made in clarifying the role of TRAF6 in mammals, the function of TRAF6 in fish is still poorly understood. In this study, we cloned the orange-spotted grouper (Epinephelus coioides) TRAF6 (EcTRAF6) cDNA, with an open reading frame of 1713bp encoding 570 amino acids. Sequence analysis indicated that EcTRAF6 contains the four characteristic domains conserved in the TRAF family, including an N-terminal RING finger, two zinc fingers, a coiled-coil domain, and a C-terminal MATH domain. Homology alignment and phylogenetic analysis demonstrated that EcTRAF6 shares high sequence identity with TRAF6 of other fish species. The EcTRAF6 gene contains seven exons and six introns, which is similar to the organization in ayu, but not in the common carp, human, or mouse (six exons and five introns). EcTRAF6 transcripts were broadly expressed in all tissues tested, and increased after infection with Cryptocaryon irritans. Intracellular localization showed EcTRAF6 was distributed mainly in the cytoplasm. Over-expression of wild type (WT) EcTRAF6, truncated forms of EcTRAF6, including ΔZinc finger 2 and ΔMATH, and a mutant of C78A activated NF-κB strongly in HEK293T cells; whereas truncations, including ΔRING, ΔZinc finger 1 and Δcoiled-coiled, and a mutant of K132R induced the activity of NF-κB slightly compared to WT EcTRAF6, implying the latter has a more crucial role in downstream signal transduction. Together, these results suggested EcTRAF6 functions like that of mammals to activate NF-κB, and it might have an important role in host defense against parasitic infections.

  5. Characterization and functional analysis of a tandem-repeat galectin-9 in large yellow croaker Larimichthys crocea.

    PubMed

    Zhang, Dong Ling; Lv, Chang Huan; Yu, Da Hui; Wang, Zhi Yong

    2016-05-01

    Galectins are a family of endogenous lectins with β-galactosides affinity, playing significant roles in the innate immunity of vertebrates and invertebrates. In this report, a new galectin-9 cDNA was identified and characterized in large yellow croaker Larimichthys crocea (designated as LcGal-9). The complete cDNA sequence of LcGal-9 was 1795 bp, with an open reading frame (ORF) of 1032 bp encoding 343 amino acids. The putative LcGal-9 protein contained two carbohydrate recognition domains (CRDs) connected by a linker peptide, with each carrying two conserved β-galactoside binding motifs H-NPR and WG-EE-, and it possessed neither a signal peptide nor a transmembrane domain. LcGal-9 protein shared 43-74% identity with galectin-9 sequences from other species. The qRT-PCR analysis revealed that LcGal-9 mRNA was constitutively expressed in all tissues examined, predominately expressed in liver, spleen, gill, kidney, head-kidney and intestine. Western blot analysis showed that LcGal-9 protein was highly expressed in liver, spleen, intestine, kidney, head-kidney, skin, gill, and heart, but not detected in muscle and plasma. LcGal-9 mRNA transcripts were induced by poly I:C in the liver (from 6 h to 48 h), spleen (at 12 h) and head-kidney (at 12 h and 24 h). In contrast, Vibrio parahaemolyticus caused a significant down-regulation in these three tissues, except for in spleen of 48 h and head-kidney of 3 h. Post-infection with Cryptocaryon irritans, the transcripts were dramatically up-regulated in gill, skin, spleen and head-kidney during initial infection period, while significant down-regulation afterward was also observed both in spleen and head-kidney. The recombinant LcGal-9 (named as rLcGal-9) purified from Escherichia coli BL21 (DE3) demonstrated hemagglutination against human, rabbit and L. crocea in a Ca(2+)-independent manner, which was inhibited by α-Lactose and LPS. The results of bacterial agglutination assays showed that rLcGal-9 was able to agglutinate

  6. Population model for Amyloodinium ocellatum infecting the spotted seatrout Cynoscion nebulosus and the red snapper Lutjanus campechanus.

    PubMed

    Masson, Ignacio; Lotz, Jeffrey M; Blaylock, Reginald B

    2013-10-11

    The dinoflagellate Amyloodinium ocellatum, a major pathogen in warm water mariculture, has a trophont, a tomont and a dinospore life history stage. This paper presents a population model for A. ocellatum infecting spotted seatrout Cynoscion nebulosus and red snapper Lutjanus campechanus and evaluates the relative effect of each vital rate on the A. ocellatum population growth rate. The vital rates were estimated by incubating trophonts in vitro and tracking their development through the successive life history stages at 25°C and 33 ppt. The A. ocellatum population growth rate was 1.90 d-1 for spotted seatrout and 1.92 d-1 for red snapper. Highest elasticity values (0.24 and 0.23 in spotted seatrout and red snapper, respectively) corresponded to transitions from the dinospore to the trophont stage, the trophont stage to the tomont stage and the tomont stage back to the dinospore stage in both host species (self-loops not included). A 50% change in vital rates showed that the mean number of dinospores produced by a tomont had the largest effect on the A. ocellatum population growth rate (15%), followed by the dinospore infection rate (14%), the tomont sporulation rate (12%) and the dinospore mortality rate (10%) in both host species. A comparison of modeled and experimental vital rate threshold values revealed a 2.5- (spotted seatrout) or a 2.6-fold (red snapper) difference in the values for dinospore mortality, which is the smallest difference among all the modeled and experimental vital rates. Therefore, measures that increase dinospore mortality have a greater likelihood of influencing the outcome of an epidemic. PMID:24113247

  7. Population model for Amyloodinium ocellatum infecting the spotted seatrout Cynoscion nebulosus and the red snapper Lutjanus campechanus.

    PubMed

    Masson, Ignacio; Lotz, Jeffrey M; Blaylock, Reginald B

    2013-10-11

    The dinoflagellate Amyloodinium ocellatum, a major pathogen in warm water mariculture, has a trophont, a tomont and a dinospore life history stage. This paper presents a population model for A. ocellatum infecting spotted seatrout Cynoscion nebulosus and red snapper Lutjanus campechanus and evaluates the relative effect of each vital rate on the A. ocellatum population growth rate. The vital rates were estimated by incubating trophonts in vitro and tracking their development through the successive life history stages at 25°C and 33 ppt. The A. ocellatum population growth rate was 1.90 d-1 for spotted seatrout and 1.92 d-1 for red snapper. Highest elasticity values (0.24 and 0.23 in spotted seatrout and red snapper, respectively) corresponded to transitions from the dinospore to the trophont stage, the trophont stage to the tomont stage and the tomont stage back to the dinospore stage in both host species (self-loops not included). A 50% change in vital rates showed that the mean number of dinospores produced by a tomont had the largest effect on the A. ocellatum population growth rate (15%), followed by the dinospore infection rate (14%), the tomont sporulation rate (12%) and the dinospore mortality rate (10%) in both host species. A comparison of modeled and experimental vital rate threshold values revealed a 2.5- (spotted seatrout) or a 2.6-fold (red snapper) difference in the values for dinospore mortality, which is the smallest difference among all the modeled and experimental vital rates. Therefore, measures that increase dinospore mortality have a greater likelihood of influencing the outcome of an epidemic.

  8. Survival and fate of Salmonella enterica serovar Montevideo in adult Horn Flies (Diptera: Muscidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Contamination of cattle peripheral lymph nodes with Salmonella enterica is proposed to occur via a transdermal route of entry. If so, bacteria may be introduced to cattle by biting arthropods. Biting flies, such as horn flies (Haematobia irritans irritans (L.); Diptera: Muscidae), are intriguing ca...

  9. Importance of supercooling points in the overwintering of the horn fly and stable fly (Diptera:Muscidae).

    PubMed

    Jones, S R; Kunz, S E

    1997-07-01

    Supercooling points were determined for eggs, 3rd instars, pupae, newly emerged unfed adults and 3-d-old engorged laboratory reared adults of Haematobia irritans (L.) and Stomoxys calcitrans (L.). Wild nondiapausing and diapausing pupae of H. irritans also were tested. Mean supercooling points ranged from -28.0 degrees C for H. irritans eggs to -6.8 degrees C for H. irritans larvae. Mean supercooling points of all H. irritans developmental stages were lower than those of comparable S. calcitrans developmental stages, with the exception of larvae where the reverse was true. The mean supercooling point of diapausing H. irritans pupae (-23.5 degrees C) was significantly lower than those of nondiapausing laboratory pupae (-20.8 degrees C) or nondiapausing wild pupae (-20.2 degrees C). Developmental stages of both species were freeze intolerant, with no survival following exposures to temperatures below the supercooling points. Results are discussed with respect to the disparate overwintering strategies of these species and in relation to typical climatic minima experienced in south central Texas. The cold tolerance of H. irritans and S. calcitrans pupae was compared at 4 degrees C, a temperature below their developmental threshold of 11.5 degrees C and above their mean supercooling points. The survival of H. irritans pupae was significantly greater than the survival of S. calcitrans pupae. Cold injury was a significant mortality factor for both species.

  10. Multiplex Polymerase Chain Reaction in the Horn Fly: Detection of Pyrethroid, Organophosphate and Cyclodiene Target Site Resistance

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The horn fly, Haematobia irritans irritans, is an important pest to the livestock industry that causes economic losses of approximately US$1 billion in the U.S. and a similar value in Latin America. Horn fly control efforts still relies mainly on direct application of insecticides although horn fly ...

  11. In vitro and in vivo assessment of the effect of antiprotozoal compounds isolated from Psoralea corylifolia against Ichthyophthirius multifiliis in fish

    PubMed Central

    Song, Kaige; Ling, Fei; Huang, Aiguo; Dong, Wenjing; Liu, Guanglu; Jiang, Chao; Zhang, Qizhong; Wang, Gaoxue

    2015-01-01

    Ichthyophthirius multifiliis, an external fish parasite, often causes significant economic damage to the aquaculture industry. Since the use of malachite green was banned, the search of alternative substance to control I. multifiliis infections becomes stringent. In present study, in vitro and in vivo anti-ich efficacies of isopsoralen and psoralidin, two active compounds isolated from methanol extract of Psoralea corylifolia by bioassay-guided fractionation based on the efficacy of anti-ich encysted tomonts, were evaluated. In vitro antiprotozoal efficacy of psoralidin is much better than that of isopsoralen. Psoralidin can kill all theronts at concentrations of 0.8 mg/L or more during 4 h exposure; and terminate reproduction of I. multifiliis post 6 h exposure of protomonts to 0.9 mg/L and encysted tomonts to 1.2 mg/L. In vivo trials showed that 5 h exposure of infected fish to 2.5 mg/L of psoralidin significantly reduced the number of theronts released from tomonts. Furthermore, we observed that a part of protomonts, collected from infected fish post treatment, presented characteristic morphological changes of apoptosis after staining with Annexin V-EGFP/propidium iodide, indicating the possible mechanism of psoralidin against I. multifiliis trophont in situ. On the basis of these results, psoralidin can be used as a potential lead compound for the development of commercial drug against I. multifiliis. PMID:26042195

  12. A Natural Cattle Immune Response Against Horn Fly (Diptera: Muscidae) Salivary Antigens May Regulate Parasite Blood Intake.

    PubMed

    Breijo, M; Pastro, L; Rocha, S; Ures, X; Alonzo, P; Santos, M; Bolatto, C; Fernández, C; Meikle, A

    2016-08-01

    The horn fly, Haematobia irritans (L.), is a blood-sucking ectoparasite that is responsible for sizeable economic losses in livestock. The salivary gland products facilitate blood intake. Taking advantage of the identification of novel H. irritans salivary antigens (Hematobin, HTB and Irritans 5, IT5), we investigated the parasite loads, H. irritans blood intake, and antibody response of naturally infected bovines during the fly season. Fly loads and fly hemoglobin content fluctuated during the trial. Each time horn fly loads exceeded 200 flies per cattle, a reduction in horn fly blood intake was observed three weeks later. All of the cattle elicited an antibody response against HTB and IT5 that declined once the fly season was over. Cattle anti-IT5 titers were positively correlated with parasite loads and negatively correlated with fly blood intake. These results suggest that the natural changes in the H. irritans blood intake observed in this study were associated with a natural host response against horn fly salivary antigens.

  13. Ectoparasites of the swift fox in northwestern Texas.

    PubMed

    Pence, Danny B; Kamler, Jan F; Ballard, Warren B

    2004-07-01

    Ectoparasites were collected from chemically immobilized swift foxes (Vulpes velox) in the Texas Panhandle (USA). Three species of fleas (Pulex irritans, Dactylopsylla percernis, and Euhoplopsyllus affinis) and one species of tick (Ixodes sculptus) were found. Pulex irritans was the only abundant ectoparasite; it occurred on all 23 foxes brushed in 1999-2000 and all but one of 34 hosts examined in 2000-01. Otherwise, this swift fox population had a depauperate ectoparasite fauna; the remainder of the ectoparasites only occurred on a few (< or =5%) of the hosts. Because of previous taxonomic confusion between P. irritans and the closely related P. simulans, the zoogeographic distribution of these two species in many areas of western North America needs to be verified. Apparently, only the human flea P. irritans occurs on wild canids in the Texas Panhandle. However, there are previous records of P. simulans on other carnivores in western and central Texas. Some of these specimens were reexamined, and their identifications were reconfirmed. Also, the recent literature on the controversial taxonomic status of these two flea species is reviewed. The male terminal aedeagal sexual apparatus is the only means currently available to separate P. irritans and P. simulans.

  14. Plague and the Human Flea, Tanzania

    PubMed Central

    Leirs, Herwig; Makundi, Rhodes H.; Van Dongen, Stefan; Davis, Stephen; Neerinckx, Simon; Deckers, Jozef; Libois, Roland

    2007-01-01

    Domestic fleas were collected in 12 villages in the western Usambara Mountains in Tanzania. Of these, 7 are considered villages with high plague frequency, where human plague was recorded during at least 6 of the 17 plague seasons between 1986 and 2004. In the remaining 5 villages with low plague frequency, plague was either rare or unrecorded. Pulex irritans, known as the human flea, was the predominant flea species (72.4%) in houses. The density of P. irritans, but not of other domestic fleas, was significantly higher in villages with a higher plague frequency or incidence. Moreover, the P. irritans index was strongly positively correlated with plague frequency and with the logarithmically transformed plague incidence. These observations suggest that in Lushoto District human fleas may play a role in plague epidemiology. These findings are of immediate public health relevance because they provide an indicator that can be surveyed to assess the risk for plague. PMID:17553245

  15. Ophryoglena hemophaga n. sp. (Ciliophora: Ophryoglenidae): a parasite of the digestive gland of zebra mussels Dreissena polymorpha.

    PubMed

    Molloy, Daniel P; Lynn, Denis H; Giamberini, Laure

    2005-07-18

    Ophryoglena hemophaga n. sp. is described from a freshwater Dreissena polymorpha population in the Rhine delta of the Netherlands. This is the first ophryoglenine species (order Hymenostomatida, suborder Ophryoglenina) recorded as a molluscan parasite. As is typical of ciliates in the suborder Ophryoglenina, O. hemophaga exhibits a polymorphic life history with cystment and reproduction by palintomy. Trophonts were observed within digestive gland lumina, and zebra mussel hemocytes were present in some of their digestive vacuoles. The presence of a single, longitudinal tract of multiple contractile vacuoles represents its most unique feature and distinguishes it from all other described Ophryoglena spp. The number of somatic kineties of O. hemophaga (range 100 to 124) is also distinguishing as it is one of the lowest for [corrected] an Ophryoglena sp. Other characteristics of this species include: ovoid to elongate trophonts 96 to 288 microm in length, with an elongate macronucleus 41 to 65 microm in length; tomonts 50 to 150 microm in diameter producing a clear mucous cyst envelope, whose thickness is approximately half of the tomont diameter; elongated theronts 96 to 131 microm in length which emerge after 1 to 3 cell divisions taking 36 to 48 h at 20 +/- 3 degrees C. Protomonts and theronts are, respectively, negatively and positively phototactic--characteristics that likely aid in maintenance of infection in zebra mussel populations.

  16. 21 CFR 524.814 - Eprinomectin.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ..., FEEDS, AND RELATED PRODUCTS OPHTHALMIC AND TOPICAL DOSAGE FORM NEW ANIMAL DRUGS § 524.814 Eprinomectin... per kilogram). (2) Indications for use. The drug is used in beef and dairy cattle for treatment and... of D. vivaparus for 21 days after treatment and H. irritans for 7 days after treatment....

  17. Salivary Gland Thrombostasin Isoforms Differentially Regulate Blood Uptake of Horn Flies Fed on Control- and Thrombostasin-Vaccinated Cattle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Thrombostasin (TS) is an anticlotting protein found in saliva of Haematobia irritans (horn flies). The polymorphic nature of the ts gene was first associated with success of horn flies blood feeding on a laboratory host, New Zealand White rabbits. In this study, we report results of similar studies ...

  18. Seasonal Variation and Frequency Distribution of Ectoparasites in Crossbreed Cattle in Southeastern Brazil

    PubMed Central

    Ferraz da Costa, Maria do Socorro; Guimarães, Marcos Pezzi; Lima, Walter dos Santos; Ferraz da Costa, Ana Julia; Facury Filho, Elias Jorge; Araujo, Ricardo Nascimento

    2014-01-01

    The aims of this study were to evaluate the seasonal variation and frequency distribution of Rhipicephalus (Boophilus) microplus, Haematobia irritans, and Dermatobia hominis on crossbred heifers under field conditions in the northeast of Minas Gerais state, southeastern Brazil. From November 2007 to September 2009 (23 months), 40 heifers aged 16.6 ± 2.4 months were divided into groups A (1/4 Holstein × 3/4 Gir) and B (1/2 Holstein × 1/2 Gir) and had the monthly infestation estimated along with the climatic conditions. The mean maximum and minimum temperatures were 28.5 and 19°C, respectively. The ectoparasites were present on animals in all months of the year. The levels of ticks on the animals were low (3.0 ± 0.2 ticks/animal), with the highest density in midwinter. The temperature was the climatic factor that most influenced the tick levels. The population of H. irritans (13.9 ± 0.3 flies/animal) and D. hominis (1.5 ± 0.2 larvae/animal) on heifers was more influenced by rainfall and exhibited two population peaks during the year. 1/2 Holstein heifers harbored significantly more H. irritans and D. hominis than 1/4 Holstein heifers. The results are discussed considering the most appropriate periods to apply ectoparasiticides and the genetic make-up of the animals. PMID:26464941

  19. Faunal distribution of fleas and their blood-feeding preferences using enzyme-linked immunosorbent assays from farm animals and human shelters in a new rural region of southern Iran.

    PubMed

    Moemenbellah-Fard, Mohammad Djaefar; Shahriari, Bahador; Azizi, Kourosh; Fakoorziba, Mohammad Reza; Mohammadi, Jalal; Amin, Masoume

    2016-03-01

    Blood sucking insects, such as fleas, are responsible for the transmission of many infectious disease-causing agents which impose an intolerable burden on the health of people living particularly in endemic parts of the world. Fleas (Insecta: Siphonaptera) are found in many parts of the world including Iran. Both adult male and female fleas are obligatory ectoparasites. They are one of the main public health concerns as a result of their nuisance or the potential to act as vectors of a number of medically-important pathogens. The current study was conducted to examine the geographical distribution and fauna of fleas and their anthropophagic index in part of Fars province, southern Iran. This study was the first to be done in Iran. A total of 20 villages were randomly selected. From October 2011 to May 2012, adult fleas were collected by direct hand catch from human to animal shelters. Overall 848 fleas, most of which were blood-fed, were captured from the floor or the body of farm animal hosts (cattle, sheep, goat and hens). Only two different genera of fleas were identified, the main species (99.76 %) was human flea, Pulex irritans. The village of Shamsabad was the most heavily infested area. P. irritans had an anthropophagic index of 15 % using indirect enzyme-linked immunosorbent assays (ELISA). It could be concluded that P. irritans is widely distributed in this area. Based on their blood feeding activity, fleas thus posed a serious health threat to residents and their economically important livestock in this part of Iran.

  20. Faunal distribution of fleas and their blood-feeding preferences using enzyme-linked immunosorbent assays from farm animals and human shelters in a new rural region of southern Iran.

    PubMed

    Moemenbellah-Fard, Mohammad Djaefar; Shahriari, Bahador; Azizi, Kourosh; Fakoorziba, Mohammad Reza; Mohammadi, Jalal; Amin, Masoume

    2016-03-01

    Blood sucking insects, such as fleas, are responsible for the transmission of many infectious disease-causing agents which impose an intolerable burden on the health of people living particularly in endemic parts of the world. Fleas (Insecta: Siphonaptera) are found in many parts of the world including Iran. Both adult male and female fleas are obligatory ectoparasites. They are one of the main public health concerns as a result of their nuisance or the potential to act as vectors of a number of medically-important pathogens. The current study was conducted to examine the geographical distribution and fauna of fleas and their anthropophagic index in part of Fars province, southern Iran. This study was the first to be done in Iran. A total of 20 villages were randomly selected. From October 2011 to May 2012, adult fleas were collected by direct hand catch from human to animal shelters. Overall 848 fleas, most of which were blood-fed, were captured from the floor or the body of farm animal hosts (cattle, sheep, goat and hens). Only two different genera of fleas were identified, the main species (99.76 %) was human flea, Pulex irritans. The village of Shamsabad was the most heavily infested area. P. irritans had an anthropophagic index of 15 % using indirect enzyme-linked immunosorbent assays (ELISA). It could be concluded that P. irritans is widely distributed in this area. Based on their blood feeding activity, fleas thus posed a serious health threat to residents and their economically important livestock in this part of Iran. PMID:27065620

  1. Porpostoma guamensis n. sp., a philasterine scuticociliate associated with brown-band disease of corals.

    PubMed

    Lobban, Christopher S; Raymundo, Laurie M; Montagnes, David J S

    2011-01-01

    Brown band disease of coral is caused by a ciliate that consumes the tissue of the corals in the genus Acropora. We describe the ciliate associated with this disease on Guam, based on: general morphology, division stages, and ciliature observed on live and protargol-stained specimens; modification of the oral structures between divisional stages, observed on protargol-stained specimens; and some aspects of behavior in field and laboratory studies. Porpostoma guamensis n. sp. is elongate and has ciliature typical for the genus; live cells are 70-500 × 20-75 μm; the macronucleus is sausage-like, elongate but often bent, positioned centrally along the main cell axis; the oral ciliature follows a basic pattern, being composed of three adoral polykinetidal regions, as described for other species in the genus, although there is variability in the organization, especially in large cells where the three regions are not easily distinguished. Ciliates fed on coral with their oral region adjacent to the tissue, which they engulfed, leaving the coral a bare skeleton. Both zooxanthellae and nematocysts from coral occurred in the ciliates. Zooxanthellae appeared to be ingested alive but deteriorated within 2-3 days. Ciliates formed thin-walled division cysts on the coral and divided up to 3 times. Cysts formed around daughter cells within cysts. We provide some observations on the complex division pattern of the ciliate (i.e. tomont-trophont-cyst) and propose a possible complete pattern that requires further validation.

  2. Porpostoma guamensis n. sp., a philasterine scuticociliate associated with brown-band disease of corals.

    PubMed

    Lobban, Christopher S; Raymundo, Laurie M; Montagnes, David J S

    2011-01-01

    Brown band disease of coral is caused by a ciliate that consumes the tissue of the corals in the genus Acropora. We describe the ciliate associated with this disease on Guam, based on: general morphology, division stages, and ciliature observed on live and protargol-stained specimens; modification of the oral structures between divisional stages, observed on protargol-stained specimens; and some aspects of behavior in field and laboratory studies. Porpostoma guamensis n. sp. is elongate and has ciliature typical for the genus; live cells are 70-500 × 20-75 μm; the macronucleus is sausage-like, elongate but often bent, positioned centrally along the main cell axis; the oral ciliature follows a basic pattern, being composed of three adoral polykinetidal regions, as described for other species in the genus, although there is variability in the organization, especially in large cells where the three regions are not easily distinguished. Ciliates fed on coral with their oral region adjacent to the tissue, which they engulfed, leaving the coral a bare skeleton. Both zooxanthellae and nematocysts from coral occurred in the ciliates. Zooxanthellae appeared to be ingested alive but deteriorated within 2-3 days. Ciliates formed thin-walled division cysts on the coral and divided up to 3 times. Cysts formed around daughter cells within cysts. We provide some observations on the complex division pattern of the ciliate (i.e. tomont-trophont-cyst) and propose a possible complete pattern that requires further validation. PMID:21205062

  3. Host site of activity and cytological effects of histone-like proteins on the parasitic dinoflagellate Amyloodinium ocellatum.

    PubMed

    Noga, Edward J; Fan, Zhiqin; Silphaduang, Umaporn

    2002-12-10

    Histone-like proteins (HLPs) are broad-spectrum, endogenously produced antibiotics which we have isolated from tissues of rainbow trout Oncorhynchus mykiss and hybrid striped bass (Morone saxatilis male x M. chrysops female). Here, we show that HLP-1, which has high sequence homology to histone H2B, equally inhibited both young and mature trophonts of the important ectoparasite Amyloodinium ocellatum. In addition to direct killing of Amyloodinium trophonts, there was evidence that HLP-1 from both rainbow trout and hybrid striped bass caused severe developmental abnormalities, including delayed development, in both the parasitic trophont stage as well as the reproductive tomont stage. The deleterious effects of HLP-1 also were manifested in what appeared to be 'delayed mortality', where parasites of normal appearance would die later in development. Similar serious damage was also seen with calf histone H2B and the unrelated peptide antibiotic magainin 2. A comparison of the antibiotic activity in mucus versus epidermis compartments of the skin of hybrid striped bass suggested that the majority of antibiotic (including HLP-1) activity resided in the epidermis, although some activity was present in the mucus. These data suggest that normal, nonimmune fish skin contains potent defenses against protozoan ectoparasites and that the effects of these defenses may extend beyond their transient interactions with the parasites, which has important implications for this host-parasite relationship.

  4. Efficacy of Brahman breeding in the management of insecticide-resistant horn flies (Diptera: Muscidae) on beef cattle.

    PubMed

    Steelman, C D; McNew, R W; Brown, M A; Tolley, G; Phillips, J M

    1994-02-01

    The efficacy of Brahman breeding used as an alternative tactic to manage insecticide-resistant populations of adult horn flies, Haematobia irritans irritans (L.), was determined. Concentration-mortality bioassays done at Booneville and Hope, AR, in 1988 and 1989, respectively, showed that horn fly populations were resistant to diazinon, pirimiphos methyl, tetrachlorvinphos, and methoxychlor. Data showed loss of field efficacy for coumaphos and delnav. Mean horn fly counts on Braham cows were significantly lower than on Angus cows for all sampling dates in 1989 and 1990. Mean fly counts on Brahman x Angus cows were approximately intermediate to the two purebred mean fly counts. Brahman breeding caused significant reductions in the number of organophosphate-resistant horn flies, which had been equal to or greater than that obtained from continued spraying with organophosphate insecticides. The Brahman x Hereford cows, which have one-eighth greater Brahman breeding than the Brangus cows, had fewer horn flies on 48 of 56 sampling dates in 1988-1990 and significantly fewer flies on 37 sampling dates. The effectiveness of Brahman breeding in causing lower numbers of insecticide-resistant horn flies significantly increased as the percentage of Brahman breeding increased. PMID:8144749

  5. Perkinsus marinus in pleasure oyster Crassostrea corteziensis from Nayarit, Pacific coast of México.

    PubMed

    Cáceres-Martínez, J; Vásquez-Yeomans, R; Padilla-Lardizábal, G; del Río Portilla, M A

    2008-09-01

    Culture of the pleasure oyster Crassostrea corteziensis is emerging as an alternative to the Pacific oyster (Crassostrea gigas) for oyster producers, who face severe mortalities since 1997 in Northwest México. For determining the health status of this species, we conducted a histopathological analysis of cultured populations from two estuaries in the Pacific coast of México. Macroscopical analysis revealed animals with transparent and retracted mantle. Histopathological analysis of these specimens showed tissue alterations and parasitic forms consistent with Perkinsus sp. infection. Stages of the parasite identified included tomont and trophozoites with an eccentric vacuole characteristic of Perkinsus spp. Pieces of tissues of infected oysters were incubated in Fluid Thioglycollate Medium (FTM) resulting in blue-black hypnospores after incubation. The identity of the parasite was confirmed by species specific PCR-based assay in DNA samples from oysters, tissue fractions from FTM cultures, and deparaffined samples with Perkinsus-like parasite detected by histology. Sequencing of positive amplified fragments (307bp) showed a sequence similar to Perkinsus marinus strain TXsc NTS ribosomal RNA gene (100% coverage and 98% identity, GenBank Accession No. AF497479.1) and to P. marinus, Genomic DNA, (100% coverage and 97% identity, GenBank Accession No. S78416.1). The prevalence of P. marinus varied from 1 to 5% in Boca del Camichín and from 1 to 6% in Pozo Chino. In general, the intensity of infection was moderate. The infection was observed in oysters from 31 to 110mm of shell length. This is the first record of P. marinus in oysters from the North America Pacific coast and the first record in C. corteziensis. The origin of this parasite in the area is unknown, but it may be associated to introductions of Crassostrea virginica from the East coast of United States of America or Gulf of México.

  6. Molecular Detection of Rickettsia felis and Bartonella henselae in Dog and Cat Fleas in Central Oromia, Ethiopia

    PubMed Central

    Kumsa, Bersissa; Parola, Philippe; Raoult, Didier; Socolovschi, Cristina

    2014-01-01

    Fleas are important vectors of several Rickettsia and Bartonella spp. that cause emerging zoonotic diseases worldwide. In this study, 303 fleas collected from domestic dogs and cats in Ethiopia and identified morphologically as Ctenocephalides felis felis, C. canis, Pulex irritans, and Echidnophaga gallinacea were tested for Rickettsia and Bartonella DNA by using molecular methods. Rickettsia felis was detected in 21% of fleas, primarily C. felis, with a similar prevalence in fleas from dogs and cats. A larger proportion of flea-infested dogs (69%) than cats (37%) harbored at least one C. felis infected with R. felis. Rickettsia typhi was not detected. Bartonella henselae DNA was detected in 6% (2 of 34) of C. felis collected from cats. Our study highlights the likelihood of human exposure to R. felis, an emerging agent of spotted fever, and B. henselae, the agent of cat-scratch disease, in urban areas in Ethiopia. PMID:24445204

  7. Fleas infesting pets in the era of emerging extra-intestinal nematodes

    PubMed Central

    2013-01-01

    Modifications in climatic conditions, movements of hosts and goods, changes in animal phenology and human behaviour and increase of wildlife, are presently concurring in the geographic spread of vectors and cardio-respiratory nematodes, e.g. Dirofilaria immitis, Angiostrongylus vasorum, Aelurostrongylus abstrusus and Capillaria aerophila. All these factors may also influence dispersion and clinical significance of fleas, thus posing relevant challenges in those regions where other parasites are emerging at the same time. Ctenocephalides felis, Ctenocephalides canis and Pulex irritans cause discomfort, nuisance, allergic reactions, anaemia, and may transmit several pathogens, some of them are of importance for public health. The present article reviews the importance of fleas in small animal practice and their sanitary relevance for dogs, cats and humans, and discusses current control methods in the present era of emerging extra-intestinal nematodes, towards a possible changing perspective for controlling key parasites affecting companion animals. PMID:23497511

  8. Development of a novel self-medicating applicator for control of internal and external parasites of wild and domestic animals.

    PubMed

    Burridge, M J; Simmons, L A; Ahrens, E H; Naudé, S A; Malan, F S

    2004-03-01

    Four trials, three in the United States and one in South Africa, were conducted to evaluate the potential value of a novel self-medicating applicator in the passive control of gastrointestinal nematodes in cattle and deer, and of files and ticks on cattle using oil-based treatments. The results of the trials demonstrated that this applicator is an effective and practical device for the passive treatment of both deer and cattle for trichostrongyle infections using the endectocide, moxidectin (Cydectin. Fort Dodge Animal Health, USA), of cattle for hom fly (Haemotobia irritans) infestations using the insecticide, cyfluthrin (CyLence, Bayer AG, Germany) and of cattle for tick infestations (in particular Amblyomma hebraeum and Rhipicephalus appendiculatus) using the acaricides deltamethrin and amitraz (Delete All, Intervet, South Africa). PMID:15185574

  9. On Some Issues of Human-Animal Studies: An Introduction.

    PubMed

    Métraux, Alexandre

    2016-03-01

    Animals are "in" - since prehistoric times when humans (or their ancient ancestors) were hunting animals, and when they fabricated the Paleolithic dog as well as the Paleolithic cat. In less general terms, animals are "in" since they received names and were listed, observed, mummified, turned into totems, and, later on, dissected, tortured under laboratory conditions, trained as experimental subjects or "purified" as model organisms. And they are massively "in" again, but now from overtly legal and moral points of view, at least since the last two decades of the twentieth century. This is to say that modern members of the species Homo sapiens have always been connected to animals of the most various kinds - from the human flea (Pulex irritans) and the cat flea (Ctenocephalides felis) to marine mammals, such as dolphins and whales, from horses to parrots, from scallops to worms, and so on.

  10. Development of a novel self-medicating applicator for control of internal and external parasites of wild and domestic animals.

    PubMed

    Burridge, M J; Simmons, L A; Ahrens, E H; Naudé, S A; Malan, F S

    2004-03-01

    Four trials, three in the United States and one in South Africa, were conducted to evaluate the potential value of a novel self-medicating applicator in the passive control of gastrointestinal nematodes in cattle and deer, and of files and ticks on cattle using oil-based treatments. The results of the trials demonstrated that this applicator is an effective and practical device for the passive treatment of both deer and cattle for trichostrongyle infections using the endectocide, moxidectin (Cydectin. Fort Dodge Animal Health, USA), of cattle for hom fly (Haemotobia irritans) infestations using the insecticide, cyfluthrin (CyLence, Bayer AG, Germany) and of cattle for tick infestations (in particular Amblyomma hebraeum and Rhipicephalus appendiculatus) using the acaricides deltamethrin and amitraz (Delete All, Intervet, South Africa).

  11. Molecular detection of Rickettsia felis and Bartonella henselae in dog and cat fleas in Central Oromia, Ethiopia.

    PubMed

    Kumsa, Bersissa; Parola, Philippe; Raoult, Didier; Socolovschi, Cristina

    2014-03-01

    Fleas are important vectors of several Rickettsia and Bartonella spp. that cause emerging zoonotic diseases worldwide. In this study, 303 fleas collected from domestic dogs and cats in Ethiopia and identified morphologically as Ctenocephalides felis felis, C. canis, Pulex irritans, and Echidnophaga gallinacea were tested for Rickettsia and Bartonella DNA by using molecular methods. Rickettsia felis was detected in 21% of fleas, primarily C. felis, with a similar prevalence in fleas from dogs and cats. A larger proportion of flea-infested dogs (69%) than cats (37%) harbored at least one C. felis infected with R. felis. Rickettsia typhi was not detected. Bartonella henselae DNA was detected in 6% (2 of 34) of C. felis collected from cats. Our study highlights the likelihood of human exposure to R. felis, an emerging agent of spotted fever, and B. henselae, the agent of cat-scratch disease, in urban areas in Ethiopia.

  12. Insecticide resistance in the horn fly: alternative control strategies.

    PubMed

    Oyarzún, M P; Quiroz, A; Birkett, M A

    2008-09-01

    The horn fly, Haematobia irritans (Linnaeus 1758) (Diptera: Muscidae) is one of the most widespread and economically important pests of cattle. Although insecticides have been used for fly control, success has been limited because of the development of insecticide resistance in all countries where the horn fly is found. This problem, along with public pressure for insecticide-free food and the prohibitive cost of developing new classes of compounds, has driven the investigation of alternative control methods that minimize or avoid the use of insecticides. This review provides details of the economic impact of horn flies, existing insecticides used for horn fly control and resistance mechanisms. Current research on new methods of horn fly control based on resistant cattle selection, semiochemicals, biological control and vaccines is also discussed.

  13. Prevalence of ectoparasitic arthropods on wild animals and cattle in the Las Merindades area (Burgos, Spain)

    PubMed Central

    Domínguez-Peñafiel, G.; Giménez-Pardo, C.; Gegúndez, M.I.; Lledó, L.

    2011-01-01

    This paper reports the prevalence of ectoparasitic arthropods in sampled groups of wild (n = 128; 16 species) and domestic (n = 69; 3 species) animals in the Las Merindades area of the Province of Burgos, Spain. The study revealed that wild animals were more infested and with a wider variety of ectoparasites than domestic animals. The parasitic prevalence was 67% for wild animals and 48% for livestock. In this way, 39% of animals were infected by ticks. Ixodes ricinus and Ixodes hexagonus were the most prevalent species whereas Dermacentor reticulatus showed affinity for the fox and wolf. The overall prevalence of parasitisation by fleas was 27%. Ctenophthalmus spp. showed the wider range host in wild animals, while Pulex irritans was the most frequent specie found. The parasitic prevalences by lice (Trichodectes melis, Trichodectes canis and Trichodectes mustelae) and by mite (Neotrombicula spp., Laelaps agilis and Sarcoptes scabiei) were 4% and 12%, respectively. In both cases only wild animals were found parasited. PMID:21894267

  14. On Some Issues of Human-Animal Studies: An Introduction.

    PubMed

    Métraux, Alexandre

    2016-03-01

    Animals are "in" - since prehistoric times when humans (or their ancient ancestors) were hunting animals, and when they fabricated the Paleolithic dog as well as the Paleolithic cat. In less general terms, animals are "in" since they received names and were listed, observed, mummified, turned into totems, and, later on, dissected, tortured under laboratory conditions, trained as experimental subjects or "purified" as model organisms. And they are massively "in" again, but now from overtly legal and moral points of view, at least since the last two decades of the twentieth century. This is to say that modern members of the species Homo sapiens have always been connected to animals of the most various kinds - from the human flea (Pulex irritans) and the cat flea (Ctenocephalides felis) to marine mammals, such as dolphins and whales, from horses to parrots, from scallops to worms, and so on. PMID:26903370

  15. Sequence change and phylogenetic signal in muscoid COII DNA sequences.

    PubMed

    Szalanski, Allen L; Owens, Carrie B

    2003-08-01

    The complete DNA sequence of the mtDNA cytochrome oxidase II gene from house fly, Musca domestica, face fly, Musca autumnalis, stable fly, Stomoxys calcitrans, horn fly, Haematobia irritans, and black garbage fly, Hydrotaea aenescens, are reported. The nucleotide sequence codes for a 229 amino acid peptide. The COII sequence is A + T rich (74.1%), with up to 12.3% nucleotide and 8.4% amino acid divergence among the five taxa. Of the 688 nucleotides encoding for the gene, 135 nucleotide sites (19.6%) are variable, and 55 (8.0%) are phylogenetically informative. A phylogenetic analysis using three calliphorids as the outgroup taxa, indicates that the two haematophagus species, horn fly and stable fly, form a sister group.

  16. Risk assessment for parasites in cultures of Diplodus puntazzo (Sparidae) in the Western Mediterranean: prospects of cross infection with Sparus aurata.

    PubMed

    Sánchez-García, Neus; Raga, Juan Antonio; Montero, Francisco E

    2014-08-29

    The sharpsnout seabream Diplodus puntazzo is of interest in Mediterranean fish farming. Disease is an important problem because parasites can spread quickly in culture conditions and fish often develop high parasite burdens. Here we assess the risk that documented parasites pose to the sustainability of D. puntazzo farming. This study specifically considers metazoan and protist parasites recorded from wild and farmed D. puntazzo in scientific literature. Risk assessment studies involve the identification, characterization and qualitative quantification of the risk in question (parasitoses in this case) and the probability of establishment. We considered the parasite species which may be difficult to manage as a priority for research into potential management strategies. Those parasites which could be transmitted from cultures of gilthead seabream (Sparus aurata) were also included in this study. Four groups of parasites represented a risk to D. puntazzo farming, ranging from moderate to high: Ciliophora, Myxozoa, Monogenea and Copepoda. Three parasite species were considered high risk to D. puntazzo cultures: Amyloodinium sp., Cryptocaryon sp. and Enteromyxum leei. These species were responsible for high mortalities in cultures of these and other fish species. In addition Sparicotyle chrysophrii, Caligus ligusticus and Gnathia vorax entail a moderate risk to D. puntazzo Mediterranean farms. No important episodes have been related to caligids and isopods in Mediterranean sparids, nevertheless they should be properly managed to prevent future problems.

  17. Risk assessment for parasites in cultures of Diplodus puntazzo (Sparidae) in the Western Mediterranean: prospects of cross infection with Sparus aurata.

    PubMed

    Sánchez-García, Neus; Raga, Juan Antonio; Montero, Francisco E

    2014-08-29

    The sharpsnout seabream Diplodus puntazzo is of interest in Mediterranean fish farming. Disease is an important problem because parasites can spread quickly in culture conditions and fish often develop high parasite burdens. Here we assess the risk that documented parasites pose to the sustainability of D. puntazzo farming. This study specifically considers metazoan and protist parasites recorded from wild and farmed D. puntazzo in scientific literature. Risk assessment studies involve the identification, characterization and qualitative quantification of the risk in question (parasitoses in this case) and the probability of establishment. We considered the parasite species which may be difficult to manage as a priority for research into potential management strategies. Those parasites which could be transmitted from cultures of gilthead seabream (Sparus aurata) were also included in this study. Four groups of parasites represented a risk to D. puntazzo farming, ranging from moderate to high: Ciliophora, Myxozoa, Monogenea and Copepoda. Three parasite species were considered high risk to D. puntazzo cultures: Amyloodinium sp., Cryptocaryon sp. and Enteromyxum leei. These species were responsible for high mortalities in cultures of these and other fish species. In addition Sparicotyle chrysophrii, Caligus ligusticus and Gnathia vorax entail a moderate risk to D. puntazzo Mediterranean farms. No important episodes have been related to caligids and isopods in Mediterranean sparids, nevertheless they should be properly managed to prevent future problems. PMID:24893693

  18. Survival and fate of Salmonella enterica serovar Montevideo in adult horn flies (Diptera: Muscidae).

    PubMed

    Olafson, Pia Untalan; Lohmeyer, Kimberly H; Edrington, Thomas S; Loneragan, Guy H

    2014-09-01

    Contamination of cattle peripheral lymph nodes with Salmonella enterica is proposed to occur via a transdermal route of entry. If so, bacteria may be introduced to cattle by biting arthropods. Biting flies, such as horn flies (Haematobia irritans irritans (L.)) (Diptera: Muscidae), are intriguing candidates for transmitting Salmonella to cattle because they provide a route of entry when they breach the skin barrier during blood feeding. Using a green fluorescent protein-expressing strain of Salmonella Montevideo (S. Montevideo-GFP), the current study demonstrated that horn fly grooming subsequent to tactile exposure to the bacteria resulted in acquisition of the bacteria on mouthparts as well as microbial ingestion. Consumption of a bloodmeal containing approximately 10(2), approximately 10(4), or 10(6) S. Montevideo-GFP resulted in horn fly colonization for up to 72 h postingestion (PI). Epifluorescent microscopy indicated that the bacteria were not localized to the crop but were observed within the endoperitrophic space, suggesting that regurgitation is not a primary route of transmission. S. Montevideo-GFP were cultured from excreta of 100% of flies beginning 6-7 h PI of a medium or high dose meal and > 12 h PI in excreta from 60% of flies fed the low-dose meal. Animal hides and manure pats are sources for horn flies to acquire the Salmonella and mechanically transmit them to an animal while feeding. Mean quantities of 5.65-67.5 x 10(2) CFU per fly were cultured from fly excreta passed within 1 d after feeding, suggesting the excreta can provide an additional microbial source on the animal's hide. PMID:25276929

  19. Survival and fate of Salmonella enterica serovar Montevideo in adult horn flies (Diptera: Muscidae).

    PubMed

    Olafson, Pia Untalan; Lohmeyer, Kimberly H; Edrington, Thomas S; Loneragan, Guy H

    2014-09-01

    Contamination of cattle peripheral lymph nodes with Salmonella enterica is proposed to occur via a transdermal route of entry. If so, bacteria may be introduced to cattle by biting arthropods. Biting flies, such as horn flies (Haematobia irritans irritans (L.)) (Diptera: Muscidae), are intriguing candidates for transmitting Salmonella to cattle because they provide a route of entry when they breach the skin barrier during blood feeding. Using a green fluorescent protein-expressing strain of Salmonella Montevideo (S. Montevideo-GFP), the current study demonstrated that horn fly grooming subsequent to tactile exposure to the bacteria resulted in acquisition of the bacteria on mouthparts as well as microbial ingestion. Consumption of a bloodmeal containing approximately 10(2), approximately 10(4), or 10(6) S. Montevideo-GFP resulted in horn fly colonization for up to 72 h postingestion (PI). Epifluorescent microscopy indicated that the bacteria were not localized to the crop but were observed within the endoperitrophic space, suggesting that regurgitation is not a primary route of transmission. S. Montevideo-GFP were cultured from excreta of 100% of flies beginning 6-7 h PI of a medium or high dose meal and > 12 h PI in excreta from 60% of flies fed the low-dose meal. Animal hides and manure pats are sources for horn flies to acquire the Salmonella and mechanically transmit them to an animal while feeding. Mean quantities of 5.65-67.5 x 10(2) CFU per fly were cultured from fly excreta passed within 1 d after feeding, suggesting the excreta can provide an additional microbial source on the animal's hide.

  20. Molecular characterization of mariner-like elements in emerald ash borer, Agrilus planipennis (Coleoptera, Polyphaga).

    PubMed

    Rivera-Vega, L; Mittapalli, O

    2010-08-01

    Emerald ash borer (EAB, Agrilus planipennis), an exotic invasive pest, has killed millions of ash trees (Fraxinus spp.) in North America and continues to threaten the very survival of the entire Fraxinus genus. Despite its high-impact status, to date very little knowledge exists for this devastating insect pest at the molecular level. Mariner-like elements (MLEs) are transposable elements, which are ubiquitous in occurrence in insects and other invertebrates. Because of their low specificity and broad host range, they can be used for epitope-tagging, gene mapping, and in vitro mutagenesis. The majority of the known MLEs are inactive due to in-frame shifts and stop codons within the open reading frame (ORF). We report on the cloning and characterization of two MLEs in A. planipennis genome (Apmar1 and Apmar2). Southern analysis indicated a very high copy number for Apmar1 and a moderate copy number for Apmar2. Phylogenetic analysis revealed that both elements belong to the irritans subfamily. Based on the high copy number for Apmar1, the full-length sequence was obtained using degenerate primers designed to the inverted terminal repeat (ITR) sequences of irritans MLEs. The recovered nucleotide sequence for Apmar1 consisted of 1,292 bases with perfect ITRs, and an ORF of 1,050 bases encoding a putative transposase of 349 amino acids. The deduced amino acid sequence of Apmar1 contained the conserved regions of mariner transposases including WVPHEL and YSPDLAP, and the D,D(34)D motif. Both Apmar1 and Apmar2 could represent useful genetic tools and provide insights on EAB adaptation.

  1. Molecular detection of Rickettsia felis in different flea species from Caldas, Colombia.

    PubMed

    Ramírez-Hernández, Alejandro; Montoya, Viviana; Martínez, Alejandra; Pérez, Jorge E; Mercado, Marcela; de la Ossa, Alberto; Vélez, Carolina; Estrada, Gloria; Correa, Maria I; Duque, Laura; Ariza, Juan S; Henao, Cesar; Valbuena, Gustavo; Hidalgo, Marylin

    2013-09-01

    Rickettsioses caused by Rickettsia felis are an emergent global threat. Historically, the northern region of the province of Caldas in Colombia has reported murine typhus cases, and recently, serological studies confirmed high seroprevalence for both R. felis and R. typhi. In the present study, fleas from seven municipalities were collected from dogs, cats, and mice. DNA was extracted and amplified by polymerase chain reaction (PCR) to identify gltA, ompB, and 17kD genes. Positive samples were sequenced to identify the species of Rickettsia. Of 1,341 fleas, Ctenocephalides felis was the most prevalent (76.7%). Positive PCR results in the three genes were evidenced in C. felis (minimum infection rates; 5.3%), C. canis (9.2%), and Pulex irritans (10.0%). Basic Local Alignment Search Tool (BLAST) analyses of sequences showed high identity values (> 98%) with R. felis, and all were highly related by phylogenetic analyses. This work shows the first detection of R. felis in fleas collected from animals in Colombia.

  2. Ectoparasite load in the crested porcupine Hystrix cristata Linnaeus, 1758 in Central Italy.

    PubMed

    Mori, Emiliano; Sforzi, Andrea; Menchetti, Mattia; Mazza, Giuseppe; Lovari, Sandro; Pisanu, Benoît

    2015-06-01

    The crested porcupine Hystrix cristata is a large body-sized rodent, occurring in Europe only in the Italian Peninsula, where it may have been introduced in early Medieval times. Its parasite fauna is currently poorly known and limited to few anecdotal observations. We have analyzed the ectoparasite load of 165 crested porcupines from Tuscany and Latium (Central Italy). Both captured and road-killed individuals were checked for fleas and ticks. Overall, only 39 porcupines were infested by four species of ticks and five of fleas. Abundance of ectoparasites was higher in areas with higher habitat richness, with respect to densely wooded areas. The most frequent species was the flea Pulex irritans (25%), whose prevalence peaked in winter probably because of optimal abiotic conditions in the porcupine's den. The remaining species of both hard ticks (Rhipicephalus bursa, Pholeoixodes hexagonus, and Ixodes ventalloi) and fleas (Paraceras melis, Ctenocephalides canis, Dasypsyllus gallinulae, and Hystrichopsylla talpae), all with prevalence lower than 5%, could be due to den sharing with other vertebrates, mainly carnivores such as, e.g., red foxes and badgers. The second most prevalent species was the generalist tick Ixodes ricinus (21%). An adult male-biased parasitism for ticks has been detected, suggesting a possible role of testosterone related immune-depressive effect. The low richness in dominant ectoparasite species, built up by locally acquired generalist taxa, provides support to the allochthonous origin of this rodent in Italy. PMID:25773184

  3. The effects of the El Niño Southern Oscillation on skin and skin-related diseases: a message from the International Society of Dermatology Climate Change Task Force.

    PubMed

    Andersen, Louise K; Davis, Mark D P

    2015-12-01

    The El Niño Southern Oscillation (ENSO) is a complex climate phenomenon occurring in the Pacific Ocean at intervals of 2-7 years. The term refers to fluctuations in ocean temperatures in the tropical eastern Pacific Ocean (El Niño [the warm phase of ENSO] and La Niña [the cool phase of ENSO]) and in atmospheric pressure across the Pacific basin (Southern Oscillation). This weather pattern is attributed with causing climate change in certain parts of the world and is associated with disease outbreaks. The question of how ENSO affects skin and skin-related disease is relatively unanswered. We aimed to review the literature describing the effects of this complex weather pattern on skin. El Niño has been associated with increases in the occurrence of actinic keratosis, tinea, pityriasis versicolor, miliaria, folliculitis, rosacea, dermatitis by Paederus irritans and Paederus sabaeus, and certain vector-borne and waterborne diseases, such as dengue fever, leishmaniasis, Chagas' disease, Barmah Forest virus, and leptospirosis, and with decreases in the occurrence of dermatitis, scabies, psoriasis, and papular urticaria. La Niña has been associated with increases in the occurrence of varicella, hand, foot, and mouth disease, and Ross River virus (in certain areas), and decreases in viral warts and leishmaniasis. Reports on the effects of ENSO on skin and skin-related disease are limited, and more studies could be helpful in the future.

  4. Molecular Detection of Rickettsia felis in Different Flea Species from Caldas, Colombia

    PubMed Central

    Ramírez-Hernández, Alejandro; Montoya, Viviana; Martínez, Alejandra; Pérez, Jorge E.; Mercado, Marcela; de la Ossa, Alberto; Vélez, Carolina; Estrada, Gloria; Correa, Maria I.; Duque, Laura; Ariza, Juan S.; Henao, Cesar; Valbuena, Gustavo; Hidalgo, Marylin

    2013-01-01

    Rickettsioses caused by Rickettsia felis are an emergent global threat. Historically, the northern region of the province of Caldas in Colombia has reported murine typhus cases, and recently, serological studies confirmed high seroprevalence for both R. felis and R. typhi. In the present study, fleas from seven municipalities were collected from dogs, cats, and mice. DNA was extracted and amplified by polymerase chain reaction (PCR) to identify gltA, ompB, and 17kD genes. Positive samples were sequenced to identify the species of Rickettsia. Of 1,341 fleas, Ctenocephalides felis was the most prevalent (76.7%). Positive PCR results in the three genes were evidenced in C. felis (minimum infection rates; 5.3%), C. canis (9.2%), and Pulex irritans (10.0%). Basic Local Alignment Search Tool (BLAST) analyses of sequences showed high identity values (> 98%) with R. felis, and all were highly related by phylogenetic analyses. This work shows the first detection of R. felis in fleas collected from animals in Colombia. PMID:23878183

  5. Fungi associated with black mould on baobab trees in southern Africa.

    PubMed

    Cruywagen, Elsie M; Crous, Pedro W; Roux, Jolanda; Slippers, Bernard; Wingfield, Michael J

    2015-07-01

    There have been numerous reports in the scientific and popular literature suggesting that African baobab (Adansonia digitata) trees are dying, with symptoms including a black mould on their bark. The aim of this study was to determine the identity of the fungi causing this black mould and to consider whether they might be affecting the health of trees. The fungi were identified by sequencing directly from mycelium on the infected tissue as well as from cultures on agar. Sequence data for the ITS region of the rDNA resulted in the identification of four fungi including Aureobasidium pullulans, Toxicocladosporium irritans and a new species of Rachicladosporium described here as Rachicladosporium africanum. A single isolate of an unknown Cladosporium sp. was also found. These fungi, referred to here as black mould, are not true sooty mould fungi and they were shown to penetrate below the bark of infected tissue, causing a distinct host reaction. Although infections can lead to dieback of small twigs on severely infected branches, the mould was not found to kill trees.

  6. Role of insects in the transmission of bovine leukosis virus: potential for transmission by stable flies, horn flies, and tabanids.

    PubMed

    Buxton, B A; Hinkle, N C; Schultz, R D

    1985-01-01

    The ability of stable flies (Stomoxys calcitrans), horn flies (Haematobia irritans), and tabanids (Diptera: Tabanidae) to transmit bovine leukosis virus (BLV) was investigated. Stable flies and horn flies were fed on blood collected from an infected cow, and the flies' mouthparts were immediately removed, placed in RPMI-1640 medium, ground, and inoculated into sheep and calves. Infection of sheep occurred with mouthparts from as few as 25 stable flies or 25 horn flies. However, sheep were not infected when removal of stable fly mouthparts was delayed greater than or equal to 1 hour after blood feeding. Infection of calves occurred after inoculation of mouthparts removed immediately after feeding from as few as 50 stable flies or 100 horn flies. Infected blood, applied by capillary action to the mouthparts (labella) of 15 deer flies (Chrysops sp) and a single horse fly (Tabanus atratus) caused infection in each of 2 sheep. Infection did not occur in 2 calves inoculated daily for 5 days with mouthparts from 50 horn flies collected after feeding on a BLV-infected steer. Four calves receiving bites from 75 stable flies interrupted from feeding on a BLV-positive cow also were not infected. Seronegative cattle held for 1 to 4 months in a screened enclosure with positive cattle in the presence of biting flies were not infected with BLV. The feeding behavior of each insect is discussed to assess their potential as vectors of BLV. PMID:2982293

  7. Wild Red Foxes (Vulpes vulpes) as Sentinels of Parasitic Diseases in the Province of Soria, Northern Spain.

    PubMed

    Lledó, Lourdes; Giménez-Pardo, Consuelo; Saz, José Vicente; Serrano, José Luis

    2015-12-01

    Four hundred red foxes (Vulpes vulpes) were examined for ecto- (arthropods) and endoparasites (Leishmania spp., Trichinella spp., and intestinal parasites). Different species of flea (total prevalence, 40.50%), tick (16.25%), mite (7.25%), and fly (1.50%) were identified. The most prevalent flea was Pulex irritans (found on 29% of the foxes); the most prevalent tick, mite, and fly were Ixodes canisuga (on 5%), Sarcoptes scabiei (on 5.25%), and Hippobosca equina (on 1%), respectively. The endoparasites identified included Leishmania spp. (found in 12% of the foxes), Trichinella spp. (in 15.5%, with T. britovi the most prevalent species in 15.25%), Cestoda (in 72.75%, with Mesocestoides spp. the most prevalent in 69.50%), and intestinal ascarids (in 73.25%, with Ancylostoma caninum the most prevalent in 12.50%). No animal was free of parasites. The present results suggest that foxes can act as sentinels of diseases transmitted by ecto- and endoparasites.

  8. Ectoparasite load in the crested porcupine Hystrix cristata Linnaeus, 1758 in Central Italy.

    PubMed

    Mori, Emiliano; Sforzi, Andrea; Menchetti, Mattia; Mazza, Giuseppe; Lovari, Sandro; Pisanu, Benoît

    2015-06-01

    The crested porcupine Hystrix cristata is a large body-sized rodent, occurring in Europe only in the Italian Peninsula, where it may have been introduced in early Medieval times. Its parasite fauna is currently poorly known and limited to few anecdotal observations. We have analyzed the ectoparasite load of 165 crested porcupines from Tuscany and Latium (Central Italy). Both captured and road-killed individuals were checked for fleas and ticks. Overall, only 39 porcupines were infested by four species of ticks and five of fleas. Abundance of ectoparasites was higher in areas with higher habitat richness, with respect to densely wooded areas. The most frequent species was the flea Pulex irritans (25%), whose prevalence peaked in winter probably because of optimal abiotic conditions in the porcupine's den. The remaining species of both hard ticks (Rhipicephalus bursa, Pholeoixodes hexagonus, and Ixodes ventalloi) and fleas (Paraceras melis, Ctenocephalides canis, Dasypsyllus gallinulae, and Hystrichopsylla talpae), all with prevalence lower than 5%, could be due to den sharing with other vertebrates, mainly carnivores such as, e.g., red foxes and badgers. The second most prevalent species was the generalist tick Ixodes ricinus (21%). An adult male-biased parasitism for ticks has been detected, suggesting a possible role of testosterone related immune-depressive effect. The low richness in dominant ectoparasite species, built up by locally acquired generalist taxa, provides support to the allochthonous origin of this rodent in Italy.

  9. The effects of the El Niño Southern Oscillation on skin and skin-related diseases: a message from the International Society of Dermatology Climate Change Task Force.

    PubMed

    Andersen, Louise K; Davis, Mark D P

    2015-12-01

    The El Niño Southern Oscillation (ENSO) is a complex climate phenomenon occurring in the Pacific Ocean at intervals of 2-7 years. The term refers to fluctuations in ocean temperatures in the tropical eastern Pacific Ocean (El Niño [the warm phase of ENSO] and La Niña [the cool phase of ENSO]) and in atmospheric pressure across the Pacific basin (Southern Oscillation). This weather pattern is attributed with causing climate change in certain parts of the world and is associated with disease outbreaks. The question of how ENSO affects skin and skin-related disease is relatively unanswered. We aimed to review the literature describing the effects of this complex weather pattern on skin. El Niño has been associated with increases in the occurrence of actinic keratosis, tinea, pityriasis versicolor, miliaria, folliculitis, rosacea, dermatitis by Paederus irritans and Paederus sabaeus, and certain vector-borne and waterborne diseases, such as dengue fever, leishmaniasis, Chagas' disease, Barmah Forest virus, and leptospirosis, and with decreases in the occurrence of dermatitis, scabies, psoriasis, and papular urticaria. La Niña has been associated with increases in the occurrence of varicella, hand, foot, and mouth disease, and Ross River virus (in certain areas), and decreases in viral warts and leishmaniasis. Reports on the effects of ENSO on skin and skin-related disease are limited, and more studies could be helpful in the future. PMID:26471012

  10. Efficacy of novaluron as a feed-through for control of immature horn flies, house flies, and stable flies (Diptera: Muscidae) developing in cow manure.

    PubMed

    Lohmeyer, K H; Pound, J M; Yeater, K M; May, M A

    2014-07-01

    Two rates (0.4 mg/kg body weight/d and 0.6 mg/kg body weight/d) of a daily feed-through formulation of novaluron (Novaluron 0.67% active ingredient Cattle Mix), a newer benzoylphenyl urea insecticide, were evaluated for efficacy in controlling the larval stage of horn flies, Haematobia irritans (L.), house flies, Musca domestica L., and stable flies, Stomoxys calcitrans (L.), developing in cow manure. Both rates of feed-through novaluron, delivered consecutively for 10 d, reduced adult emergence of all three species when compared with the untreated control. The presence of deformed puparia indicated that novaluron had an insect growth regulator effect on the developing fly larvae. Both of the feed-through rates evaluated resulted in 100% reduction of adult stable fly emergence after the second day of feed-through treatment. The level of control efficacy observed against these three fly species make this feed-through formulation a candidate for use in an integrated livestock pest management program, particularly in confined cattle production situations where a feed-through product could be easily administered.

  11. Fleas of the San Joaquin kit fox (Vulpes velox macrotis) on Camp Roberts Army National Guard Training Site, California

    SciTech Connect

    Spencer, K.A.; Egoscue, H.J.

    1992-09-01

    A total of 3,241 fleas, representing seven species, were identified from 398 samples collected from San Joaquin kit foxes (Vulpes velox macrotis), California ground squirrels (Spermophilus beecheyi), and deer mice (Peromyscus maniculatus) at Camp Roberts Army National Guard Training Site, California, from November 1988 through September 1991. Of 3,109 fleas collected from kit foxes 95.7% were Echidnophaga gallinacea, 4.0% Pulex irritans, 0.2% Hoplopsyllus anomolus, and 0.1% Odontopsyllus dentatus. One male Ctenocephalides fells was also collected from a kit fox. The 118 fleas collected from California ground squirrels consisted of Hoplopsyllus anomolus (55.9%), Echidnophaga gallinacea (37.3%), and Oropsylla montanus (6.8%). The 14 fleas collected from deer mice were Aetheca wagneri. Based on the distribution and abundance of flea species collected, and the vector efficiency of these fleas, it appears that kit foxes could play a role in the transfer of natural vectors of sylvatic plague between rodent populations, if the bacterium responsible for plague (Yersinia pestis) were present at Camp Roberts. Little information regarding kit fox food habits was evidenced by the distribution and abundance of small mammal flea species collected from kit foxes.

  12. Plague in Tanzania: an overview.

    PubMed

    Ziwa, Michael H; Matee, Mecky I; Hang'ombe, Bernard M; Lyamuya, Eligius F; Kilonzo, Bukheti S

    2013-10-01

    Human plague remains a public health concern in Tanzania despite its quiescence in most foci for years, considering the recurrence nature of the disease. Despite the long-standing history of this problem, there have not been recent reviews of the current knowledge on plague in Tanzania. This work aimed at providing a current overview of plague in Tanzania in terms of its introduction, potential reservoirs, possible causes of plague persistence and repeated outbreaks in the country. Plague is believed to have been introduced to Tanzania from the Middle East through Uganda with the first authentication in 1886. Xenopsylla brasiliensis, X. cheopis, Dinopsyllus lypusus, and Pulex irritans are among potential vectors while Lophuromys spp, Praomys delectorum, Graphiurus murinus, Lemniscomys striatus, Mastomys natalensis, and Rattus rattus may be the potential reservoirs. Plague persistence and repeated outbreaks in Tanzania are likely to be attributable to a complexity of factors including cultural, socio-economical, environmental and biological. Minimizing or preventing people's proximity to rodents is probably the most effective means of preventing plague outbreaks in humans in the future. In conclusion, much has been done on plague diagnosis in Tanzania. However, in order to achieve new insights into the features of plague epidemiology in the country, and to reorganize an effective control strategy, we recommend broader studies that will include the ecology of the pathogen, vectors and potential hosts, identifying the reservoirs, dynamics of infection and landscape ecology.

  13. Effect of aggregation of horn fly populations within cattle herds and consequences for sampling to obtain unbiased estimates of abundance.

    PubMed

    Lysyk, T J; Steelman, C D

    2004-07-01

    Reanalysis of counts of horn fly, Hematobia irritans (L.), obtained from a variety of cattle herds indicated that aggregation of the flies within herds decreased as mean fly density increased. Aggregation was also related to the proportion of fly-resistant and fly-susceptible cattle in a herd. Herds were grouped according to their degree of horn fly aggregation. Low aggregation herds included larger framed Angus, Horned Hereford, Polled Hereford, and Red Poll breeds. Moderate aggregation occurred with Brahman, Charolais, small-framed Angus, mixed cows, and Hereford x Charolais cross. High aggregation occurred with Chianina and mixed herds. Relationships between the sample means and variances varied among aggregation groups. A resampling approach was used to determine the influence of random sampling of a herd on the proportion of horn fly population estimates within fixed percentages of the true mean. The proportion of sample means within +/- 5, 10, 15, and 20% of the true means varied with the proportion of the herd sampled, the mean and variance of fly density, and herd size. Recommendations for obtaining sample size to estimate fly density within a fixed percentage of the true mean are given.

  14. Detection of target site resistance to pyrethroids and organophosphates in the horn fly using multiplex polymerase chain reaction.

    PubMed

    Foil, L D; Guerrero, F D; Bendele, K G

    2010-09-01

    The horn fly, Haematobia irritans L., is an obligate blood-feeding fly and the primary insect pest parasitizing cattle in the United States. Pesticide resistance has become a substantial problem for cattle producers, and although several mechanisms of resistance are possible, target site resistance is the most important mechanism preventing control of this fly in the United States and possibly other countries. We developed a multiplex polymerase chain reaction assay to detect the known target site, pyrethroid resistance-associated mutation in the horn fly and a recently reported G262A mutation in the horn fly acetylcholinesterase, the target site for organophosphates. As expected, the pyrethroid resistance target site mutation was found in fly populations from Texas, Louisiana, Washington, Georgia, Mexico, and Brazil. This mutation was found to have a gender bias as it was more prevalent in females than males. The G262A acetylcholinesterase mutation was found in Texas, Louisiana, Washington, Georgia, and Mexico, but not Brazil. There was no gender bias in the occurrence of this mutation, and there was no correlation between the occurrence of the kdr and the G262A mutations. Unlike the case with the pyrethroid target site mutation, the presence of G262A did not appear to exclusively provide the level of resistance required to account for bioassay results. It is likely an additional mutation(s) occurs in the target site and/or a metabolic resistance mechanism exists in organophosphate-resistant horn fly populations.

  15. [The man in the ice under special consideration of paleo-pathological evidence].

    PubMed

    Spindler, K

    2001-01-01

    Dealing with archaeological findings, the physical condition of the 5000-year-old Man in the Ice will be discussed. Several research projects with autoscopical, radiological, computertomographical and laparoscopical methods present an overview of the state of health of the Tyrolean Iceman. The vascular network at the lower brain shows a slight hardening of the arteries. On the right side of the thorax the sixth and seventh ribs are fractured without callus formation. On the left side serial rib fractures can be seen which had healed well. The twelfth ribs are absent on both sides. The Iceman was suffering from arthrosis of the right hip of medium severity. Osteolysis of the little toe is indicative of frostbite. Some ecto- and endoparasites were found (Pulex irritans, Trichuris trichiura). The Iceman has a number of skin markings on various places. The location of these tattoos matches the X-ray findings of discrete to medium arthrotic changes in the corresponding joints. It seems reasonable to assume that the markings were applied for a therapeutic reason. The combination of thickening of the arteries, arthrosis of the joints, the healed fractures and frostbite lesions illustrates the hard life to be endured by the Man in the Ice and by Late Stone Age man in general.

  16. Wild Red Foxes (Vulpes vulpes) as Sentinels of Parasitic Diseases in the Province of Soria, Northern Spain.

    PubMed

    Lledó, Lourdes; Giménez-Pardo, Consuelo; Saz, José Vicente; Serrano, José Luis

    2015-12-01

    Four hundred red foxes (Vulpes vulpes) were examined for ecto- (arthropods) and endoparasites (Leishmania spp., Trichinella spp., and intestinal parasites). Different species of flea (total prevalence, 40.50%), tick (16.25%), mite (7.25%), and fly (1.50%) were identified. The most prevalent flea was Pulex irritans (found on 29% of the foxes); the most prevalent tick, mite, and fly were Ixodes canisuga (on 5%), Sarcoptes scabiei (on 5.25%), and Hippobosca equina (on 1%), respectively. The endoparasites identified included Leishmania spp. (found in 12% of the foxes), Trichinella spp. (in 15.5%, with T. britovi the most prevalent species in 15.25%), Cestoda (in 72.75%, with Mesocestoides spp. the most prevalent in 69.50%), and intestinal ascarids (in 73.25%, with Ancylostoma caninum the most prevalent in 12.50%). No animal was free of parasites. The present results suggest that foxes can act as sentinels of diseases transmitted by ecto- and endoparasites. PMID:26565688

  17. Experimental infection of Columbian black-tailed deer with the Lyme disease spirochete.

    PubMed

    Lane, R S; Berger, D M; Casher, L E; Burgdorfer, W

    1994-01-01

    The course of Borrelia burgdorferi-infection in Columbian black-tailed deer. (Odocoileus hemionus columbianus), its effect on the health of these animals, and their reservoir competence for fleas were evaluated experimentally. Four yearling females inoculated intramuscularly with 10(8) organisms of the CA4 strain of B. burgdorferi, and two yearling males unexposed to spirochetes, were monitored daily for 3 mo. Spirochetes were reisolated from the blood of three does at 14 or 70 days postinjection, and from several tissues of the fourth doe at necropsy. Considerable antigenic heterogeneity was observed among the reisolates as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Only two of the four infected deer developed significant antibodies (> or = 1:128) to B. burgdorferi with titers persisting for < or = 2 mo. Hematological values were highly variable and the degree of variation observed was much greater than that reported previously for Columbian black-tailed deer or other subspecies of mule deer. Infected deer did not manifest signs of Lyme disease. On histologic examination of eight tissues per deer, we observed a minimal hepatic lesion in all animals exposed to B. burgdorferi. No spirochetes were detected in 367 fleas (Pulex irritans) that had naturally infested these deer; thus this flea probably is an inefficient host of B. burgdorferi.

  18. In vivo transposon mutagenesis of the methanogenic archaeon Methanosarcina acetivorans C2A using a modified version of the insect mariner-family transposable element Himar1

    PubMed Central

    Zhang, J. K.; Pritchett, M. A.; Lampe, D. J.; Robertson, H. M.; Metcalf, W. W.

    2000-01-01

    We present here a method for in vivo transposon mutagenesis of a methanogenic archaeon, Methanosarcina acetivorans C2A, which because of its independence from host-specific factors may have broad application among many microorganisms. Because there are no known Methanosarcina transposons we modified the mariner transposable element Himar1, originally found in the insect Hematobia irritans, to allow its use in this organism. This element was chosen because, like other mariner elements, its transposition is independent of host factors, requiring only its cognate transposase. Modified mini-Himar1 elements were constructed that carry selectable markers that are functional in Methanosarcina species and that express the Himar1 transposase from known Methanosarcina promoters. These mini-mariner elements transpose at high frequency in M. acetivorans to random sites in the genome. The presence of an Escherichia coli selectable marker and plasmid origin of replication within the mini-mariner elements allows facile cloning of these transposon insertions to identify the mutated gene. In preliminary experiments, we have isolated numerous mini-mariner-induced M. acetivorans mutants, including ones with insertions that confer resistance to toxic analogs and in genes that encode proteins involved in heat shock, nitrogen fixation, and cell-wall structures. PMID:10920201

  19. Parasites of importance for human health in Nigerian dogs: high prevalence and limited knowledge of pet owners

    PubMed Central

    Ugbomoiko, Uade Samuel; Ariza, Liana; Heukelbach, Jorg

    2008-01-01

    Background Dogs are the most common pet animals worldwide. They may harbour a wide range of parasites with zoonotic potential, thus causing a health risk to humans. In Nigeria, epidemiological knowledge on these parasites is limited. Methods In a community-based study, we examined 396 dogs in urban and rural areas of Ilorin (Kwara State, Central Nigeria) for ectoparasites and intestinal helminths. In addition, a questionnaire regarding knowledge and practices was applied to pet owners. Results Nine ectoparasite species belonging to four taxa and six intestinal helminth species were identified: fleas (Ctenocephalides canis, Pulex irritans, Tunga penetrans), mites (Demodex canis, Otodectes sp., Sarcoptes scabiei var. canis), ticks (Rhipicephalus sanguineus, Ixodes sp.), and lice (Trichodectes canis); and Toxocara canis, Ancylostoma sp., Trichuris vulpis, Dipylidium caninum, Taenidae and Strongyloides sp. Overall prevalence of ectoparasites was 60.4% and of intestinal helminths 68.4%. The occurrence of C. canis, R. sanguineus, T. canis, Ancylostoma sp. and T. vulpis was most common (prevalence 14.4% to 41.7%). Prevalence patterns in helminths were age-dependent, with T. canis showing a decreasing prevalence with age of host, and a reverse trend in other parasite species. Knowledge regarding zoonoses was very limited and the diseases not considered a major health problem. Treatment with antiparasitic drugs was more frequent in urban areas. Conclusion Parasites of importance for human health were highly prevalent in Nigerian dogs. Interventions should include health education provided to dog owners and the establishment of a program focusing on zoonotic diseases. PMID:19068110

  20. Effect of aggregation of horn fly populations within cattle herds and consequences for sampling to obtain unbiased estimates of abundance.

    PubMed

    Lysyk, T J; Steelman, C D

    2004-07-01

    Reanalysis of counts of horn fly, Hematobia irritans (L.), obtained from a variety of cattle herds indicated that aggregation of the flies within herds decreased as mean fly density increased. Aggregation was also related to the proportion of fly-resistant and fly-susceptible cattle in a herd. Herds were grouped according to their degree of horn fly aggregation. Low aggregation herds included larger framed Angus, Horned Hereford, Polled Hereford, and Red Poll breeds. Moderate aggregation occurred with Brahman, Charolais, small-framed Angus, mixed cows, and Hereford x Charolais cross. High aggregation occurred with Chianina and mixed herds. Relationships between the sample means and variances varied among aggregation groups. A resampling approach was used to determine the influence of random sampling of a herd on the proportion of horn fly population estimates within fixed percentages of the true mean. The proportion of sample means within +/- 5, 10, 15, and 20% of the true means varied with the proportion of the herd sampled, the mean and variance of fly density, and herd size. Recommendations for obtaining sample size to estimate fly density within a fixed percentage of the true mean are given. PMID:15311450

  1. Reassessment of the potential economic impact of cattle parasites in Brazil.

    PubMed

    Grisi, Laerte; Leite, Romário Cerqueira; Martins, João Ricardo de Souza; Barros, Antonio Thadeu Medeiros de; Andreotti, Renato; Cançado, Paulo Henrique Duarte; León, Adalberto Angel Pérez de; Pereira, Jairo Barros; Villela, Humberto Silva

    2014-01-01

    The profitability of livestock activities can be diminished significantly by the effects of parasites. Economic losses caused by cattle parasites in Brazil were estimated on an annual basis, considering the total number of animals at risk and the potential detrimental effects of parasitism on cattle productivity. Estimates in U.S. dollars (USD) were based on reported yield losses among untreated animals and reflected some of the effects of parasitic diseases. Relevant parasites that affect cattle productivity in Brazil, and their economic impact in USD billions include: gastrointestinal nematodes - $7.11; cattle tick (Rhipicephalus (Boophilus) microplus) - $3.24; horn fly (Haematobia irritans) - $2.56; cattle grub (Dermatobia hominis) - $0.38; New World screwworm fly (Cochliomyia hominivorax) - $0.34; and stable fly (Stomoxys calcitrans) - $0.34. The combined annual economic loss due to internal and external parasites of cattle in Brazil considered here was estimated to be at least USD 13.96 billion. These findings are discussed in the context of methodologies and research that are required in order to improve the accuracy of these economic impact assessments. This information needs to be taken into consideration when developing sustainable policies for mitigating the impact of parasitism on the profitability of Brazilian cattle producers.

  2. Structure and evolution of the mitochondrial genome of Exorista sorbillans: the Tachinidae (Diptera: Calyptratae) perspective.

    PubMed

    Shao, Yuan-jun; Hu, Xian-qiong; Peng, Guang-da; Wang, Rui-xian; Gao, Rui-na; Lin, Chao; Shen, Wei-de; Li, Rui; Li, Bing

    2012-12-01

    The first complete mitochondrial genome (mitogenome) of Tachinidae Exorista sorbillans (Diptera) is sequenced by PCR-based approach. The circular mitogenome is 14,960 bp long and has the representative mitochondrial gene (mt gene) organization and order of Diptera. All protein-coding sequences are initiated with ATN codon; however, the only exception is Cox I gene, which has a 4-bp ATCG putative start codon. Ten of the thirteen protein-coding genes have a complete termination codon (TAA), but the rest are seated on the H strand with incomplete codons. The mitogenome of E. sorbillans is biased toward A+T content at 78.4 %, and the strand-specific bias is in reflection of the third codon positions of mt genes, and their T/C ratios as strand indictor are higher on the H strand more than those on the L strand pointing at any strain of seven Diptera flies. The length of the A+T-rich region of E. sorbillans is 106 bp, including a tandem triple copies of a13-bp fragment. Compared to Haematobia irritans, E. sorbillans holds distant relationship with Drosophila. Phylogenetic topologies based on the amino acid sequences, supporting that E. sorbillans (Tachinidae) is clustered with strains of Calliphoridae and Oestridae, and superfamily Oestroidea are polyphyletic groups with Muscidae in a clade.

  3. Resistance of beef cattle of two genetic groups to ectoparasites and gastrointestinal nematodes in the state of São Paulo, Brazil.

    PubMed

    Oliveira, M C S; Alencar, M M; Giglioti, R; Beraldo, M C D; Aníbal, F F; Correia, R O; Boschini, L; Chagas, A C S; Bilhassi, T B; Oliveira, H N

    2013-10-18

    The resistance to infestations by ectoparasites and infections by gastrointestinal nematodes was studied in 45 animals (males and females) of two genetic groups: purebred Nelore (NI, n=28) and Three-Cross (1/2 Angus+1/4 Canchim+1/4 Nelore - TC, n=17). The animals were monitored for 24 months, during which they were left to graze in tropical pastures without receiving treatment for parasites. Each month the animals were examined for infestations by external parasites, to count the numbers of cattle ticks Rhipicephalus microplus with diameter greater than 4.5mm present on the left side, horn flies (Haematobia irritans) present in the lumbar region and botfly larvae (Dermatobia hominis) present on the entire body. The H. irritans counts were performed with the aid of digital photographs. At the time of examination, fecal samples were collected to count the eggs per gram (EPG) and to perform coprocultures, and peripheral blood samples were drawn to determine the packed cell volume (PCV) and to count the eosinophils. For statistical analysis, the count data were transformed into log₁₀ (n+1), where n is the number of parasites. For PCV, significant effects (P<0.05) were found for collection month (CO), genetic group (GG) and gender (SX), with means and respective standard errors of 41.5 ± 0.65% for the NI animals, 39.3 ± 0.83% for the TC, 41.5 ± 0.72% for the females and 39.3 ± 0.77% for the males. Regarding the eosinophil counts, only the effect of sex was significant (P<0.01), with means and respective standard errors of 926.0 ± 46.2/μL, for males and 1088.0 ± 43.8/μL of blood, for females. The NI animals presented lower mean counts for all the external parasites compared to the TC animals (P<0.01). For ticks, the transformed means followed by standard errors for the NI and TC animals were 0.06 ± 0.01 and 0.34 ± 0.02, while for horn flies these were 0.92 ± 0.05 and 1.36 ± 0.06 and for botfly larvae they were 0.05 ± 0.03 and 0.45 ± 0.05, respectively

  4. Selective inhibitors of digestive enzymes from Aedes aegypti larvae identified by phage display.

    PubMed

    Soares, Tatiane Sanches; Soares Torquato, Ricardo Jose; Alves Lemos, Francisco Jose; Tanaka, Aparecida Sadae

    2013-01-01

    Dengue is a serious disease transmitted by the mosquito Aedes aegypti during blood meal feeding. It is estimated that the dengue virus is transmitted to millions of individuals each year in tropical and subtropical areas. Dengue control strategies have been based on controlling the vector, Ae. aegypti, using insecticide, but the emergence of resistance poses new challenges. The aim of this study was the identification of specific protease inhibitors of the digestive enzymes from Ae. aegypti larvae, which may serve as a prospective alternative biocontrol method. High affinity protein inhibitors were selected by all of the digestive serine proteases of the 4th instar larval midgut, and the specificity of these inhibitors was characterized. These inhibitors were obtained from a phage library displaying variants of HiTI, a trypsin inhibitor from Haematobia irritans, that are mutated in the reactive loop (P1-P4'). Based on the selected amino acid sequence pattern, seven HiTI inhibitor variants were cloned, expressed and purified. The results indicate that the HiTI variants named T6 (RGGAV) and T128 (WNEGL) were selected by larval trypsin-like (IC(50) of 1.1 nM) and chymotrypsin-like enzymes (IC(50) of 11.6 nM), respectively. The variants T23 (LLGGL) and T149 (GGVWR) inhibited both larval chymotrypsin-like (IC(50) of 4.2 nM and 29.0 nM, respectively) and elastase-like enzymes (IC(50) of 1.2 nM for both). Specific inhibitors were successfully obtained for the digestive enzymes of Ae. aegypti larvae by phage display. Our data also strongly suggest the presence of elastase-like enzymes in Ae. aegypti larvae. The HiTI variants T6 and T23 are good candidates for the development as a larvicide to control the vector.

  5. Lousicidal, ovicidal and repellent efficacy of some essential oils against lice and flies infesting water buffaloes in Egypt.

    PubMed

    Khater, Hanem F; Ramadan, Mohamed Y; El-Madawy, Reham S

    2009-10-14

    The lousicidal and repellent effects of five essential oils were investigated for the first time against the buffalo louse, Haematopinus tuberculatus, and flies infesting water buffaloes in Qalyubia Governorate, Egypt. For the in vitro studies, filter paper contact bioassays were used to test the oils and their lethal activities were compared with that of d-phenothrin. Four minutes post-treatment, the median lethal concentration, LC50, values were 2.74, 7.28, 12.35, 18.67 and 22.79% for camphor (Cinnamomum camphora), onion (Allium cepa), peppermint (Mentha piperita), chamomile (Matricaria chamomilla) and rosemary oils (Rosmarinus officinalis), respectively, whereas for d-phenothrin, it was 1.17%. The lethal time (50) (LT50) values were 0.89, 2.75, 15.39, 21.32, 11.60 and 1.94 min after treatment with 7.5% camphor, onion, peppermint, chamomile, rosemary and d-phenothrin, respectively. All the materials used except rosemary, which was not applied, were ovicidal to the eggs of H. tuberculatus. Despite the results of the in vitro assays, the in vivo treatments revealed that the pediculicidal activity was more pronounced with oils. All treated lice were killed after 0.5-2 min, whereas with d-phenothrin, 100% mortality was reached only after 120 min. The number of lice infesting buffaloes was significantly reduced 3, 6, 4, 6 and 9 days after treatment with camphor, peppermint, chamomile, onion, and d-phenothrin, respectively. Moreover, the oils and d-phenothrin significantly repelled flies, Musca domestica, Stomoxys calcitrans, Haematobia irritans and Hippobosca equina, for 6 and 3 days post-treatment, respectively. No adverse effects were noted on either animals or pour-on operators after exposure to the applied materials. Consequently, some Egyptian essential oils show potential for the development of new, speedy and safe lousicides and insect repellents for controlling lice and flies which infest water buffaloes. PMID:19596520

  6. Control of lone star ticks (Acari: Ixodidae) on Spanish goats and white-tailed deer with orally administered ivermectin.

    PubMed

    Miller, J A; Garris, G I; George, J E; Oehler, D D

    1989-12-01

    Ivermectin administered orally to Spanish goats, Capra hircus (L.), or to white-tailed deer, Odocoileus virginianus (Zimmerman), was highly effective against lone star ticks, Amblyomma americanum (L.). For Spanish goats, daily oral doses of 20 micrograms/kg resulted in greater than or equal to 2 ppb ivermectin in the blood. This level was sufficient to cause greater than 95% reduction of estimated larvae from feeding ticks. A bioassay with horn flies, Haematobia irritans (L.), was developed to estimate oral intake of ivermectin. Probit analysis of dose-mortality data indicated that a 50% reduction in adult horn fly emergence can be expected when the manure from goats treated orally with ivermectin at 10, 20, 35, and 50 micrograms/kg/d was mixed with untreated cow manure at a rate of 0.345, 0.110, 0.100, and 0.092%, respectively. In studies with white-tailed deer, daily oral doses of 35 and 50 micrograms/kg/d provided 100% control of adult and about 90% control of nymphs that were placed on treated fawns. A single oral dose of 50 micrograms/kg gave greater than 90% control of adult and nymphal ticks attached to treated fawns at the time of drug administration and 70% control of ticks placed on treated deer three days thereafter. When ticks were placed on fawns treated with a single dose of ivermectin (50 micrograms/kg) the engorgement period was longer, ticks were lighter in weight, and females laid fewer eggs than ticks detaching from control fawns. A single oral dose of ivermectin at 20 micrograms/kg prevented about 60% of the adult and nymphal ticks attached at the time of drug administration from engorging, but did not affect other ticks placed on the animals after treatment. PMID:2607030

  7. Prevalence of ectoparasites in free-range backyard chickens, domestic pigeons (Columba livia domestica) and turkeys of Kermanshah province, west of Iran.

    PubMed

    Rezaei, Farid; Hashemnia, Mohammad; Chalechale, Abdolali; Seidi, Shahin; Gholizadeh, Maryam

    2016-06-01

    This study was carried out on free-range backyard chickens, domestic pigeons (Columba livia domestica) and turkeys from May 2012 to April 2013 to determine the prevalence and identify the species of ectoparasites in Kermanshah province, west of Iran. Of the total of 600 free-range backyard chickens (185 ♂ and 415 ♀), 700 domestic pigeons (278 ♂ and 422 ♀) and 150 turkeys (53 ♂ and 97 ♀), 389 (64.83 %), 608 (86.85 %) and 54 (36 %) were infected with one or more parasites respectively. Eleven ectoparasites species including five of lice (50.16 % Menacanthus stramineus, 13.66 % Menopon gallinae, 4.83 % Cuclotogaster heterographus, 5.16 % Goniocotes gallinae, 2.33 % Goniodes gigas), three of mites (26.33 % Dermanyssus gallinae, 8.5 % Ornithonyssus bursa, 7 % Cnemidocoptes mutans), one of tick (78.66 % Argas persicus) and two of flea (12.33 % Echidnophaga gallinacea, 2 % Pulex irritans) were found in the backyard chickens. The domestic pigeons were infected with six species of parasites including: Columbicola columbae (61.7 %), M. gallinae (10.43 %), M. stramineus (9 %), D. gallinae (8.28 %), Argas reflexus (74.14 %) and Pseudolynchia canariensis (27.7 %). The ectoparasites species recorded in turkeys were M. gallinae (14 %), M. stramineus (8 %), D. gallinae (12.66 %), C. mutans (6 %), A. persicus (24.66 %) and E. gallinacean (6 %). This is the first survey to determine the prevalence and identify the species of ectoparasites among free-range backyard chicken, domestic pigeons and turkeys in Kermanshah province. The high prevalence rate of ectoparasites in free-range backyard chickens and domestic pigeons indicates that parasitic infection is a common problem in this area.

  8. Control of lone star ticks (Acari: Ixodidae) on Spanish goats and white-tailed deer with orally administered ivermectin.

    PubMed

    Miller, J A; Garris, G I; George, J E; Oehler, D D

    1989-12-01

    Ivermectin administered orally to Spanish goats, Capra hircus (L.), or to white-tailed deer, Odocoileus virginianus (Zimmerman), was highly effective against lone star ticks, Amblyomma americanum (L.). For Spanish goats, daily oral doses of 20 micrograms/kg resulted in greater than or equal to 2 ppb ivermectin in the blood. This level was sufficient to cause greater than 95% reduction of estimated larvae from feeding ticks. A bioassay with horn flies, Haematobia irritans (L.), was developed to estimate oral intake of ivermectin. Probit analysis of dose-mortality data indicated that a 50% reduction in adult horn fly emergence can be expected when the manure from goats treated orally with ivermectin at 10, 20, 35, and 50 micrograms/kg/d was mixed with untreated cow manure at a rate of 0.345, 0.110, 0.100, and 0.092%, respectively. In studies with white-tailed deer, daily oral doses of 35 and 50 micrograms/kg/d provided 100% control of adult and about 90% control of nymphs that were placed on treated fawns. A single oral dose of 50 micrograms/kg gave greater than 90% control of adult and nymphal ticks attached to treated fawns at the time of drug administration and 70% control of ticks placed on treated deer three days thereafter. When ticks were placed on fawns treated with a single dose of ivermectin (50 micrograms/kg) the engorgement period was longer, ticks were lighter in weight, and females laid fewer eggs than ticks detaching from control fawns. A single oral dose of ivermectin at 20 micrograms/kg prevented about 60% of the adult and nymphal ticks attached at the time of drug administration from engorging, but did not affect other ticks placed on the animals after treatment.

  9. Microarray analysis of female- and larval-specific gene expression in the horn fly (Diptera: Muscidae).

    PubMed

    Guerrero, Felix D; Dowd, Scot E; Sun, Yan; Saldivar, Leonel; Wiley, Graham B; Macmil, Simone L; Najar, Fares; Roe, Bruce A; Foil, Lane D

    2009-03-01

    The horn fly, Haematobia irritans L., is an obligate blood-feeding parasite of cattle, and control of this pest is a continuing problem because the fly is becoming resistant to pesticides. Dominant conditional lethal gene systems are being studied as population control technologies against agricultural pests. One of the components of these systems is a female-specific gene promoter that drives expression of a lethality-inducing gene. To identify candidate genes to supply this promoter, microarrays were designed from a horn fly expressed sequence tag (EST) database and probed to identify female-specific and larval-specific gene expression. Analysis of dye swap experiments found 432 and 417 transcripts whose expression levels were higher or lower in adult female flies, respectively, compared with adult male flies. Additionally, 419 and 871 transcripts were identified whose expression levels were higher or lower in first-instar larvae compared with adult flies, respectively. Three transcripts were expressed more highly in adult females flies compared with adult males and also higher in the first-instar larval lifestage compared with adult flies. One of these transcripts, a putative nanos ortholog, has a high female-to-male expression ratio, a moderate expression level in first-instar larvae, and has been well characterized in Drosophila. melanogaster (Meigen). In conclusion, we used microarray technology, verified by reverse transcriptase-polymerase chain reaction and massively parallel pyrosequencing, to study life stage- and sex-specific gene expression in the horn fly and identified three gene candidates for detailed evaluation as a gene promoter source for the development of a female-specific conditional lethality system.

  10. Prevalence of Fleas and Gastrointestinal Parasites in Free-Roaming Cats in Central Mexico

    PubMed Central

    Cantó, Germinal J.; Guerrero, Roberto I.; Olvera-Ramírez, Andrea M.; Milián, Feliciano; Mosqueda, Juan; Aguilar-Tipacamú, Gabriela

    2013-01-01

    The prevalence of fleas and gastrointestinal parasites in free-roaming and domestic cats in central Mexico was evaluated. Three hundred and fifty eight cats captured in the street or brought in by owners to the Animal Control Center Unit, a unit of State Government, from June 2010 to May 2011, were included in the study. All cats were examined for the presence of fleas and gastrointestinal worms. One-hundred and ninety (53%) cats were infested with at least one flea species. Single infestations were observed in 106 (30%) cats and mixed infestations in 84 (23%) cats. Four species of fleas were recovered: Ctenocephalides felis in 53% of the cats, C. canis in 18%, Echidnophaga gallinacea in 7% and Pulex irritans in 1%. One-hundred and sixty three (45%) cats were infected with one or more species of gastrointestinal parasites: 48 (13%) with nematodes, 145 (40%) with cestodes, and one animal presented Moniliformis moniliformis. Prevalences and mean intensity of infection were: Physaloptera praeputialis 7 and 18; T. cati 3 and 2; Ancylostoma tubaeforme 2.5 and 2; Toxascaris leonina 0.5 and 2; Dipylidium caninum 36 and 32; Taenia taeniformis 4 and 3 and Moniliformis moniliformis 0.3 and 106, respectively. There was significant association (P<0.01), between season and ectoparasites load, more fleas were obtained in the summer and autumn than in the winter and spring; however, no statistical difference was observed for endoparasites load (P>0.05). The correlation between the total number of ectoparasites and endoparasites was not significant (r = 0.089, P = 0.094). PMID:23573282

  11. Lousicidal, ovicidal and repellent efficacy of some essential oils against lice and flies infesting water buffaloes in Egypt.

    PubMed

    Khater, Hanem F; Ramadan, Mohamed Y; El-Madawy, Reham S

    2009-10-14

    The lousicidal and repellent effects of five essential oils were investigated for the first time against the buffalo louse, Haematopinus tuberculatus, and flies infesting water buffaloes in Qalyubia Governorate, Egypt. For the in vitro studies, filter paper contact bioassays were used to test the oils and their lethal activities were compared with that of d-phenothrin. Four minutes post-treatment, the median lethal concentration, LC50, values were 2.74, 7.28, 12.35, 18.67 and 22.79% for camphor (Cinnamomum camphora), onion (Allium cepa), peppermint (Mentha piperita), chamomile (Matricaria chamomilla) and rosemary oils (Rosmarinus officinalis), respectively, whereas for d-phenothrin, it was 1.17%. The lethal time (50) (LT50) values were 0.89, 2.75, 15.39, 21.32, 11.60 and 1.94 min after treatment with 7.5% camphor, onion, peppermint, chamomile, rosemary and d-phenothrin, respectively. All the materials used except rosemary, which was not applied, were ovicidal to the eggs of H. tuberculatus. Despite the results of the in vitro assays, the in vivo treatments revealed that the pediculicidal activity was more pronounced with oils. All treated lice were killed after 0.5-2 min, whereas with d-phenothrin, 100% mortality was reached only after 120 min. The number of lice infesting buffaloes was significantly reduced 3, 6, 4, 6 and 9 days after treatment with camphor, peppermint, chamomile, onion, and d-phenothrin, respectively. Moreover, the oils and d-phenothrin significantly repelled flies, Musca domestica, Stomoxys calcitrans, Haematobia irritans and Hippobosca equina, for 6 and 3 days post-treatment, respectively. No adverse effects were noted on either animals or pour-on operators after exposure to the applied materials. Consequently, some Egyptian essential oils show potential for the development of new, speedy and safe lousicides and insect repellents for controlling lice and flies which infest water buffaloes.

  12. Prevalence of fleas and gastrointestinal parasites in free-roaming cats in central Mexico.

    PubMed

    Cantó, Germinal J; Guerrero, Roberto I; Olvera-Ramírez, Andrea M; Milián, Feliciano; Mosqueda, Juan; Aguilar-Tipacamú, Gabriela

    2013-01-01

    The prevalence of fleas and gastrointestinal parasites in free-roaming and domestic cats in central Mexico was evaluated. Three hundred and fifty eight cats captured in the street or brought in by owners to the Animal Control Center Unit, a unit of State Government, from June 2010 to May 2011, were included in the study. All cats were examined for the presence of fleas and gastrointestinal worms. One-hundred and ninety (53%) cats were infested with at least one flea species. Single infestations were observed in 106 (30%) cats and mixed infestations in 84 (23%) cats. Four species of fleas were recovered: Ctenocephalides felis in 53% of the cats, C. canis in 18%, Echidnophaga gallinacea in 7% and Pulex irritans in 1%. One-hundred and sixty three (45%) cats were infected with one or more species of gastrointestinal parasites: 48 (13%) with nematodes, 145 (40%) with cestodes, and one animal presented Moniliformis moniliformis. Prevalences and mean intensity of infection were: Physaloptera praeputialis 7 and 18; T. cati 3 and 2; Ancylostoma tubaeforme 2.5 and 2; Toxascaris leonina 0.5 and 2; Dipylidium caninum 36 and 32; Taenia taeniformis 4 and 3 and Moniliformis moniliformis 0.3 and 106, respectively. There was significant association (P<0.01), between season and ectoparasites load, more fleas were obtained in the summer and autumn than in the winter and spring; however, no statistical difference was observed for endoparasites load (P>0.05). The correlation between the total number of ectoparasites and endoparasites was not significant (r = 0.089, P = 0.094).

  13. Molecular detection of Rickettsia felis and Candidatus Rickettsia Asemboensis in Fleas from Human Habitats, Asembo, Kenya

    PubMed Central

    Jiang, Ju; Maina, Alice N.; Knobel, Darryn L.; Cleaveland, Sarah; Laudisoit, Anne; Wamburu, Kabura; Ogola, Eric; Parola, Philippe; Breiman, Robert F.; Njenga, M. Kariuki

    2013-01-01

    Abstract The flea-borne rickettsioses murine typhus (Rickettsia typhi) and flea-borne spotted fever (FBSF) (Rickettsia felis) are febrile diseases distributed among humans worldwide. Murine typhus has been known to be endemic to Kenya since the 1950s, but FBSF was only recently documented in northeastern (2010) and western (2012) Kenya. To characterize the potential exposure of humans in Kenya to flea-borne rickettsioses, a total of 330 fleas (134 pools) including 5 species (Xenopsylla cheopis, Ctenocephalides felis, Ctenocephalides canis, Pulex irritans, and Echidnophaga gallinacea) were collected from domestic and peridomestic animals and from human dwellings within Asembo, western Kenya. DNA was extracted from the 134 pooled flea samples and 89 (66.4%) pools tested positively for rickettsial DNA by 2 genus-specific quantitative real-time PCR (qPCR) assays based upon the citrate synthase (gltA) and 17-kD antigen genes and the Rfelis qPCR assay. Sequences from the 17-kD antigen gene, the outer membrane protein (omp)B, and 2 R. felis plasmid genes (pRF and pRFd) of 12 selected rickettsia-positive samples revealed a unique Rickettsia sp. (n=11) and R. felis (n=1). Depiction of the new rickettsia by multilocus sequence typing (MLST) targeting the 16S rRNA (rrs), 17-kD antigen gene, gltA, ompA, ompB, and surface cell antigen 4 (sca4), shows that it is most closely related to R. felis but genetically dissimilar enough to be considered a separate species provisionally named Candidatus Rickettsia asemboensis. Subsequently, 81 of the 134 (60.4%) flea pools tested positively for Candidatus Rickettsia asemboensis by a newly developed agent-specific qPCR assay, Rasemb. R. felis was identified in 9 of the 134 (6.7%) flea pools, and R. typhi the causative agent of murine typhus was not detected in any of 78 rickettsia-positive pools assessed using a species-specific qPCR assay, Rtyph. Two pools were found to contain both R. felis and Candidatus Rickettsia asemboensis DNA and 1

  14. Parasiticide resistance in flies, lice and ticks in New Zealand and Australia: mechanisms, prevalence and prevention.

    PubMed

    Heath, Acg; Levot, G W

    2015-07-01

    This review outlines the history of parasiticide resistance in the principal ectoparasites of sheep and cattle in New Zealand and Australia, blowflies, buffalo fly (Haematobia irritans exigua), sheep biting louse (Bovicola ovis) and cattle ticks, and discusses recent changes in their response to insecticides and acaricides. Mechanisms of resistance and evaluation methods are described, with ways in which insecticide resistance can be avoided or ameliorated also discussed. Resistance in sheep blowflies (Lucilia cuprina; L. sericata) to organophosphates and benzoylphenyl urea compounds is widespread in Australia and New Zealand, but there are lesser concerns about a pyrimidine carbonitrile product as well as cyromazine and macrocyclic lactone actives which still offer the promise of long-term protection. In Australia the effectiveness of synthetic pyrethroid and benzoylphenyl urea products against the biting louse of sheep have been reduced by widespread resistance but effective temephos, macrocyclic lactone, imidacloprid and spinosyn-based products are now available. Pyrethroid-resistant sheep lice are also present in New Zealand. Buffalo fly remains a significant problem in Australia and control relies heavily on insecticide use. Resistance to synthetic pyrethroids is widespread and organophosphate resistance common, but less evenly distributed. There is no evidence of resistance to acaricides used against the New Zealand cattle tick, Haemaphysalis longicornis and experience in Japan with the most commonly available active, flumethrin (a synthetic pyrethroid), suggests that this three-host tick is, in the short term, likely to remain susceptible. The same cannot be said for Rhipicephalus australis (formerly Boophilus microplus) which, in some strains, is highly resistant to many of the active ingredients in the acaricides used against it. A formamidine, a benzoylphenyl urea and macrocyclic lactones still show efficacy although some resistance is being detected to

  15. Parasites of sheep herding dogs in central Germany.

    PubMed

    Rehbein, Steffen; Kaulfuß, Karl-heinz; Visser, Martin; Sommer, Maria Franziska; Grimm, Felix; Silaghi, Cornelia

    2016-01-01

    This paper reports on endoparasite infections diagnosed in 2012 by standard coproscopical techniques and coproantigen Giardia ELISA in 165 dogs used for sheep herding in 36 farms in central Germany. The overall prevalence of dogs with evidence of endoparasite infections was 27.3% (95% CI 20.6-34.7). The most frequently identified faecal forms were those of ascarids (Toxocara, 6.7%; Toxascaris 3.6%), hookworms (5.5%) and taeniid cestodes (4.2%), followed by those of Trichuris whipworms (3.0%), Capillaria aerophila (1.8%), Angiostrongylus and Crenosoma lungworms (1.2% each) and Cystoisospora canis coccidians (0.6%). Molecular identification demonstrated the seven dogs shedding taeniid eggs positive for Taenia (T.) species tapeworms (five, T. hydatigena; one, T. ovis; one Taenia sp.). Screening of the faeces with the coproantigen ELISA revealed Giardia specific antigen in 5.5% of the samples. The majority of the dogs had evidence of single endoparasite infections (22.4%) while evidence for infection with two or three parasites concurrently was found in six (3.6%) and two (1.2%) of the dogs, respectively. Dogs ≤ 1 year (n = 19) were parasitized more frequently (p < 0.05) with overall gastrointestinal parasites (63.2% vs. 20.5%), ascarids (36.8% vs. 6.8%) and Giardia spp. (21.1% vs. 3.4%) than older dogs (n = 146). Dogs which had been wormed within six months of examination tested less frequently positive for gastrointestinal helminths compared to dogs not wormed (11.1% vs. 25.0%; p = 0.0567). In addition, ear swabs taken from 43 sheep dogs in 2012 were examined, and Otodectes cynotis mites were extracted from one dog. Identification of ectoparasites collected by full body search and combing from 113 sheep dogs in the years 2011 to 2013 revealed infestation of fleas and ticks (each up to five specimens per dog) on 13 and 108 dogs, respectively, with nine dogs carrying both fleas and ticks. Archaeopsylla erinacei, Ctenocephalides (C) canis, C. felis and Pulex irritans

  16. Detection of Rickettsia felis, Rickettsia typhi, Bartonella Species and Yersinia pestis in Fleas (Siphonaptera) from Africa

    PubMed Central

    Leulmi, Hamza; Socolovschi, Cristina; Laudisoit, Anne; Houemenou, Gualbert; Davoust, Bernard; Bitam, Idir; Raoult, Didier; Parola, Philippe

    2014-01-01

    Little is known about the presence/absence and prevalence of Rickettsia spp, Bartonella spp. and Yersinia pestis in domestic and urban flea populations in tropical and subtropical African countries. Methodology/Principal findings Fleas collected in Benin, the United Republic of Tanzania and the Democratic Republic of the Congo were investigated for the presence and identity of Rickettsia spp., Bartonella spp. and Yersinia pestis using two qPCR systems or qPCR and standard PCR. In Xenopsylla cheopis fleas collected from Cotonou (Benin), Rickettsia typhi was detected in 1% (2/199), and an uncultured Bartonella sp. was detected in 34.7% (69/199). In the Lushoto district (United Republic of Tanzania), R. typhi DNA was detected in 10% (2/20) of Xenopsylla brasiliensis, and Rickettsia felis was detected in 65% (13/20) of Ctenocephalides felis strongylus, 71.4% (5/7) of Ctenocephalides canis and 25% (5/20) of Ctenophthalmus calceatus calceatus. In the Democratic Republic of the Congo, R. felis was detected in 56.5% (13/23) of Ct. f. felis from Kinshasa, in 26.3% (10/38) of Ct. f. felis and 9% (1/11) of Leptopsylla aethiopica aethiopica from Ituri district and in 19.2% (5/26) of Ct. f. strongylus and 4.7% (1/21) of Echidnophaga gallinacea. Bartonella sp. was also detected in 36.3% (4/11) of L. a. aethiopica. Finally, in Ituri, Y. pestis DNA was detected in 3.8% (1/26) of Ct. f. strongylus and 10% (3/30) of Pulex irritans from the villages of Wanyale and Zaa. Conclusion Most flea-borne infections are neglected diseases which should be monitored systematically in domestic rural and urban human populations to assess their epidemiological and clinical relevance. Finally, the presence of Y. pestis DNA in fleas captured in households was unexpected and raises a series of questions regarding the role of free fleas in the transmission of plague in rural Africa, especially in remote areas where the flea density in houses is high. PMID:25299702

  17. Parasiticide resistance in flies, lice and ticks in New Zealand and Australia: mechanisms, prevalence and prevention.

    PubMed

    Heath, Acg; Levot, G W

    2015-07-01

    This review outlines the history of parasiticide resistance in the principal ectoparasites of sheep and cattle in New Zealand and Australia, blowflies, buffalo fly (Haematobia irritans exigua), sheep biting louse (Bovicola ovis) and cattle ticks, and discusses recent changes in their response to insecticides and acaricides. Mechanisms of resistance and evaluation methods are described, with ways in which insecticide resistance can be avoided or ameliorated also discussed. Resistance in sheep blowflies (Lucilia cuprina; L. sericata) to organophosphates and benzoylphenyl urea compounds is widespread in Australia and New Zealand, but there are lesser concerns about a pyrimidine carbonitrile product as well as cyromazine and macrocyclic lactone actives which still offer the promise of long-term protection. In Australia the effectiveness of synthetic pyrethroid and benzoylphenyl urea products against the biting louse of sheep have been reduced by widespread resistance but effective temephos, macrocyclic lactone, imidacloprid and spinosyn-based products are now available. Pyrethroid-resistant sheep lice are also present in New Zealand. Buffalo fly remains a significant problem in Australia and control relies heavily on insecticide use. Resistance to synthetic pyrethroids is widespread and organophosphate resistance common, but less evenly distributed. There is no evidence of resistance to acaricides used against the New Zealand cattle tick, Haemaphysalis longicornis and experience in Japan with the most commonly available active, flumethrin (a synthetic pyrethroid), suggests that this three-host tick is, in the short term, likely to remain susceptible. The same cannot be said for Rhipicephalus australis (formerly Boophilus microplus) which, in some strains, is highly resistant to many of the active ingredients in the acaricides used against it. A formamidine, a benzoylphenyl urea and macrocyclic lactones still show efficacy although some resistance is being detected to

  18. Parasites of sheep herding dogs in central Germany.

    PubMed

    Rehbein, Steffen; Kaulfuß, Karl-heinz; Visser, Martin; Sommer, Maria Franziska; Grimm, Felix; Silaghi, Cornelia

    2016-01-01

    This paper reports on endoparasite infections diagnosed in 2012 by standard coproscopical techniques and coproantigen Giardia ELISA in 165 dogs used for sheep herding in 36 farms in central Germany. The overall prevalence of dogs with evidence of endoparasite infections was 27.3% (95% CI 20.6-34.7). The most frequently identified faecal forms were those of ascarids (Toxocara, 6.7%; Toxascaris 3.6%), hookworms (5.5%) and taeniid cestodes (4.2%), followed by those of Trichuris whipworms (3.0%), Capillaria aerophila (1.8%), Angiostrongylus and Crenosoma lungworms (1.2% each) and Cystoisospora canis coccidians (0.6%). Molecular identification demonstrated the seven dogs shedding taeniid eggs positive for Taenia (T.) species tapeworms (five, T. hydatigena; one, T. ovis; one Taenia sp.). Screening of the faeces with the coproantigen ELISA revealed Giardia specific antigen in 5.5% of the samples. The majority of the dogs had evidence of single endoparasite infections (22.4%) while evidence for infection with two or three parasites concurrently was found in six (3.6%) and two (1.2%) of the dogs, respectively. Dogs ≤ 1 year (n = 19) were parasitized more frequently (p < 0.05) with overall gastrointestinal parasites (63.2% vs. 20.5%), ascarids (36.8% vs. 6.8%) and Giardia spp. (21.1% vs. 3.4%) than older dogs (n = 146). Dogs which had been wormed within six months of examination tested less frequently positive for gastrointestinal helminths compared to dogs not wormed (11.1% vs. 25.0%; p = 0.0567). In addition, ear swabs taken from 43 sheep dogs in 2012 were examined, and Otodectes cynotis mites were extracted from one dog. Identification of ectoparasites collected by full body search and combing from 113 sheep dogs in the years 2011 to 2013 revealed infestation of fleas and ticks (each up to five specimens per dog) on 13 and 108 dogs, respectively, with nine dogs carrying both fleas and ticks. Archaeopsylla erinacei, Ctenocephalides (C) canis, C. felis and Pulex irritans

  19. Endectocide activity of a pour-on formulation containing 1.5 per cent ivermectin +0.5 per cent abamectin in cattle

    PubMed Central

    Silva, Heloisa Cristina; Prette, Nancy; Lopes, Welber Daniel Zanetti; Sakamoto, Cláudio Alessandro M; Buzzulini, Carolina; dos Santos, Thais Rabelo; Cruz, Breno Cayeiro; Teixeira, Weslen F Pires; Felippelli, Gustavo; Carvalho, Rafael Silveira; Maciel, Willian Giquelin; Soares, Vando Edésio; da Costa, Alvimar José

    2015-01-01

    The present work aimed to evaluate, through ten different studies, the therapeutic efficacy of a new pour-on formulation, containing 1.5 per cent ivermectin +0.5 per cent abamectin, against parasites of cattle. Results obtained on trials against Rhipicephalus (Boophilus) microplus showed that the pour-on combination of 1.5 per cent ivermectin +0.5 per cent abamectin obtained superior efficacy indexes against this ectoparasite, when compared with formulations containing 0.5 per cent ivermectin, 1 per cent ivermectin and the combination of 1 per cent abamectin +20 per cent levamisole. The results of efficacy of the ivermectin+abamectin and the 0.5 per cent ivermectin against Haematobia irritans were similar. Against Cochliomyia hominivorax larvae, all pour-on formulations tested (1.5 per cent ivermectin +0.5 per cent abamectin, 0.5 per cent ivermectin and 0.5 per cent abamectin), as well as 1 per cent doramectin administered subcutaneously, were considered ineffective. Cattle medicated with 1.5 per cent ivermectin +0.5 per cent abamectin, pour-on, remained free from parasitism by Dermatobia hominis larvae during 42 days (96 per cent efficacy), while values superior to 90 per cent were obtained by 0.5 per cent ivermectin (92 per cent) and 0.5 per cent abamectin (93 per cent) until the 42nd and 35th days post treatment, respectively. Against Haemonchus placei and Oesophagostomum radiatum, the pour-on of ivermectin+abamectin showed better efficacy than the 0.5 per cent ivermectin and 0.5 per cent abamectin. As to Cooperia punctata, there was no difference regarding efficacy results obtained by the avermectins combination and abamectin. The pour-on combination of 1.5 per cent ivermectin +0.5 per cent abamectin obtained high efficacy against R. (B.) microplus, D. hominis and some species of cattle gastrointestinal helminths when compared with formulations of 0.5 per cent ivermectin and 0.5 per cent abamectin administered through the same route. PMID:26392893

  20. Molecular detection of Rickettsia felis and Candidatus Rickettsia asemboensis in fleas from human habitats, Asembo, Kenya.

    PubMed

    Jiang, Ju; Maina, Alice N; Knobel, Darryn L; Cleaveland, Sarah; Laudisoit, Anne; Wamburu, Kabura; Ogola, Eric; Parola, Philippe; Breiman, Robert F; Njenga, M Kariuki; Richards, Allen L

    2013-08-01

    The flea-borne rickettsioses murine typhus (Rickettsia typhi) and flea-borne spotted fever (FBSF) (Rickettsia felis) are febrile diseases distributed among humans worldwide. Murine typhus has been known to be endemic to Kenya since the 1950s, but FBSF was only recently documented in northeastern (2010) and western (2012) Kenya. To characterize the potential exposure of humans in Kenya to flea-borne rickettsioses, a total of 330 fleas (134 pools) including 5 species (Xenopsylla cheopis, Ctenocephalides felis, Ctenocephalides canis, Pulex irritans, and Echidnophaga gallinacea) were collected from domestic and peridomestic animals and from human dwellings within Asembo, western Kenya. DNA was extracted from the 134 pooled flea samples and 89 (66.4%) pools tested positively for rickettsial DNA by 2 genus-specific quantitative real-time PCR (qPCR) assays based upon the citrate synthase (gltA) and 17-kD antigen genes and the Rfelis qPCR assay. Sequences from the 17-kD antigen gene, the outer membrane protein (omp)B, and 2 R. felis plasmid genes (pRF and pRFd) of 12 selected rickettsia-positive samples revealed a unique Rickettsia sp. (n=11) and R. felis (n=1). Depiction of the new rickettsia by multilocus sequence typing (MLST) targeting the 16S rRNA (rrs), 17-kD antigen gene, gltA, ompA, ompB, and surface cell antigen 4 (sca4), shows that it is most closely related to R. felis but genetically dissimilar enough to be considered a separate species provisionally named Candidatus Rickettsia asemboensis. Subsequently, 81 of the 134 (60.4%) flea pools tested positively for Candidatus Rickettsia asemboensis by a newly developed agent-specific qPCR assay, Rasemb. R. felis was identified in 9 of the 134 (6.7%) flea pools, and R. typhi the causative agent of murine typhus was not detected in any of 78 rickettsia-positive pools assessed using a species-specific qPCR assay, Rtyph. Two pools were found to contain both R. felis and Candidatus Rickettsia asemboensis DNA and 1 pool