The role of apoptosis in immunosuppression of dogs with demodicosis

The aim of the present study was to evaluate the status of apoptosis in peripheral blood leukocytes of dogs with demodicosis. A total of 26 dogs suffering from demodicosis, and positive for Demodex canis mites by skin scraping, participated in the study, 13 with localized demodicosis (LD) and 13 with generalized demodicosis (GD). A further 13 clinically healthy dogs, all of whom were negative for mites upon skin scraping, were used as controls. The dogs with GD revealed significantly higher (P ≤ 0.0001) percentage of leukocytes with externalization of phosphatidylserine (PS) and depolarized mitochondrial membrane potentials (ΔΨm) as compared with the dogs with LD and healthy controls. These dogs also revealed significantly lower values (P ≤ 0.0001) of hematological parameters viz. hemoglobin, total erythrocytes count total leukocytes count, lymphocytes, monocytes and neutrophils. Significantly higher (P ≤ 0.0001) percentages of leukocytes with externalization of PS and depolarized ΔΨm were also found in dogs with LD as compared with the healthy controls. These dogs also revealed significantly lower values of Hb (P ≤ 0.0001), TEC (P=0.025), TLC (P ≤ 0.0001), lymphocytes (P=0.008), monocytes (P ≤ 0.0001) and neutrophils (P=0.03). It is concluded that premature apoptosis of PBL may be implicated in the immunosuppression of the dogs with demodicosis.     

Dietary bovine lactoferrin induces changes in immunity level and disease resistance in Asian catfish Clarias batrachus

The effects of including bovine lactoferrin (Lf) in the diet of the Asian catfish (Clarias batrachus) on specific and non-specific immunity as well as disease resistance were investigated. The catfish were fed four different diets for 2 weeks: a commercial diet as control and the same diet supplemented with 50, 100 and 200mg bovine Lf/kg feed. After 1 and 2 weeks, serum bacterial agglutination titre against Aeromonas hydrophila as a measure of specific immunity; natural serum haemolysin titre, lysozyme activity and oxidative radical production by neutrophils as a measure of non-specific immunity as well as disease resistance against A. hydrophila challenge to vaccinated and non-vaccinated animals were evaluated. The results showed that Lf supplements, particularly at 100mg level, significantly (P<0.05) enhanced serum lysozyme level, oxidative radical production and level of protection against A. hydrophila challenge in non-vaccinated animals irrespective of length of exposure. The specific immunity was not influenced by Lf feeding as evidenced from the bacterial agglutination titre and level of protection in vaccinated animals. As Lf feeding at 100mg/kg for 1 week is able to enhance the non-specific immunity and disease resistance of catfish efficiently, these results support the possible use of Lf as an immunostimulant for farmed catfish.     

Molecular characterization of <Emphasis Type="Italic">Xiphinema brevicollum</Emphasis> (Nematoda: Longidoridae) from the Czech Republic

Populations of Xiphinema brevicollum occurring in the Czech Republic were described morphologically and molecularly. Published species-specific primer set BL18 and BV3 was used to amplify three populations of X. brevicollum from the Czech Republic. These primers were tested against 9 species of Xiphinema and 11 species of Longidorus. Amplification was also observed for X. inaequale, X. italiae and X. lambertii. Three additional markers, mitochondrial cytochrome c oxidase subunit 1, ribosomal D2/D3 expansion segment of the 28S gene and 18S gene, were amplified and sequenced for X. brevicollum, X. inaequale and X. lambertii belonging to X. americanum-group. Comparison of cox1 sequences of X. brevicollum from the Czech Republic with X. taylori from the Slovak Republic (accession number AM086702) suggested that these populations represent the same species.     

Nicotinamide dehydrogenase subunit 4 analysis of Xiphinema diversicaudatum and Xiphinema simile (Nematoda: Longidoridae)

Regions within the mitochondrial gene encoding for the nicotinamide dehydrogenase subunit 4 (nad4) were characterized to evaluate the extent of genetic variation within and among Xiphinema diversicaudatum and Xiphinema simile populations. Four different sequence variants of nad4 were determined among eight populations of X. diversicaudatum and three variants among three populations of X. simile. Nucleotide variation was detected in 28 of 411 bp (2.43 to 4.87 %) in X. diversicaudatum and in three of 395 bp (0.25 to 0.76 %) in X. simile. This represents the first study based on the characterization of the nad4 gene for the analysis of population genetic of two Xiphinema species.     

Molecular characterization and diagnostics of stubby root and virus vector nematodes of the family Trichodoridae (Nematoda: Triplonchida) using ribosomal RNA genes

Plant parasitic nematodes of the family Trichodoridae cause substantial yield losses in many agricultural crops. Rapid and accurate identification of trichodorids to the species level is critical for selection of appropriate measures for control. This study analysed 99 sequences of the D2–D3 expansion segments of the 28S rRNA gene and 131 sequences of the 18S rRNA gene from the stubby nematodes belonging to the genera Nanidorus, Paratrichodorus and Trichodorus. Species delimiting was based on the integration of morphological identification, which is not provided in the present article, and molecular‐based phylogenetic inference and sequence analysis. Twenty‐two valid species and several species complexes were identified among nematodes included in the analysis. PCR‐RFLPs of the partial 18S rDNA and the D2–D3 expansion segments of the 28S rDNA were tested and proposed for identification of these nematodes. Gel PCR‐RFLP profiles and tables with restriction fragment lengths for several diagnostic enzymes are provided for identification. Some problems of taxonomy and phylogeny of nematodes of the family Trichodoridae are also discussed.     

Cloning, nucleotide sequence and phylogenetic analyses, and tissue-specific expression of the transferrin gene in Cirrhinus mrigala infected with Aeromonas hydrophila

Transferrin partial complementary DNAs were cloned from the livers of five species in four genera of Indian carps (Indian major carp species: Labeo rohita, Catla catla and Cirrhinus mrigala; medium carp: Puntius sarana; minor carp: Labeo bata) subsequent to polymerase chain reaction amplification with published heterologous primers or self-designed primers derived from conserved regions of transferrin cDNA sequences. The partial transferrin cDNAs of the five species of carps had sizes from 624 to 633 bp (487 bp for L. rohita) and encoded an open reading frame consisting of 206–211 (162 for L. rohita) amino acids. The alignments of carp cDNA sequences showed 85–97% homology and 71–93% homology in deduced amino acid sequences. A phylogenetic tree of amino acid sequences of transferrin cDNAs from carps showed that the relationship among the four genera of Indian carps is well correlated with that derived from classic morphologic analyses. The hypothesized cleavage site and interdomain bridge of transferrin molecule were predicted for the above carp species and interestingly the cleavage site amino acid sequence was found to be conserved among all the carps. To study the tissue-specific expression of the transferrin gene, various tissues (liver, kidney, spleen, brain, muscle, testis, heart, intestine, gill and fin) from apparently healthy (control), moribund and survived C. mrigala experimentally infected with Aeromonas hydrophila infection were analyzed. Transferrin mRNA was detected only in liver RNA and to lesser extent in brain tissue out of the 10 tissues analyzed irrespective of bacterial infection.     

Broad bean wilt virus: Host range,purification, serology,transmission characteristics,and occurrence in faba bean in West Asia and North Africa

A virus affecting faba bean in West Asia and Norht Africa was identified as broad bean wilt virus (BBWV) by host reactions, particle morphology and size, serology and transmission characteristics. An isolate from Syria (SV3-88) and one from Egypt (EV319-86) were found to be serologically identical and of serotype I. In host-range studies, the Syrian isolate infected systemically 59 out of 87 plant species tested. The virus was transmitted non-persistently by four aphid species naturally prevalent in Syria, but most efficiently byMyzus persicae. Inoculation of faba bean with SV3-88 14 weeks (pre-flowering) and 6 weeks after sowing (flowering) led to 25.8 and 1.8% yield loss and seed-transmission rates of 0.6 and 0.4%, respectively. The isolate SV3-88 was purified from systemically infected faba bean and yield 1.5–2 mg of partially purified virus per 100 g of leaves. When samples, with symptoms suggestive of virus infection, were collected during 1985–1989 from a number of countries in West Asia and North Africa and tested by ELISA, the virus was detected in 8 out of 127 samples tested (8/127) from Egypt, 0/44 from Lebanon, 1/23 from Morocco, 38/485 from the Sudan, 38/385 from Syria and 23/138 from Tunisia.Samenvatting Een virus uit veldboon of faba boon (Vicia faba) in West Azië en Noord-Afrika werd als tuinboneverwelkingsvirus (Fabavirus-groep) herkend aan zijn waardplantreacties, deeltjesvorm en grootte, serologie en wijze van overdracht. Een isolaat uit Syrië (SV3-88) en één uit Egypte (EV319-86) bleken serologisch identiek te zijn en te behoren tot serotype I van het virus. Met het Syrische isolaat kon in 59 van de 87 getoetste plantesoorten systemische infectie worden verkregen. Met vier veel in Syrië voorkomende bladluissorten kon het virus worden overgebracht, maar metMyzus persicae naar de meeste plantesoorten. Inoculatie van veldboon met SV3-88 vóór de bloei (14 weken na het zaaien) en tijdens de bloei (16 weken na het zaaien) gaf aanleiding tot respectivelijk 25,8 en 1,8% opbrengstreductie en tot 0,6 en 0,4% zaadoverdracht. Bij zuivering van isolaat SV3-88 uit systemisch geïnfecteerde fababoon was de opbrengst tot 1,5 à 2 mg gedeeltelijk gezuiverd virus per 100 g blad. Bij ELISA-toetsing in 1985–1989 van een groot aantal monsters afkomstig uit een aantal landen in West-Azië en Noord-Afrika werd het virus aangetoond in 8 van de 127 (8/127) monsters uit Egypte, 0/44 uit Libanon, 1/23 uit Marokko, 38/485 uit Soedan, 38/385 uit Syrië en 23/138 uit Tunesië.