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Humoral and cell-mediated immune responses of foxes (Vulpes vulpes) after experimental primary and secondary oral vaccination using SAG2 and V-RG vaccines.

Humoral and cell-mediated immune responses of 36 captive foxes to two oral vaccines against rabies currently used for foxes in Europe were studied. The Street Alabama Dufferin (SAD) mutant Gif (SAG2) vaccine has been selected by double mutation from the SAD virus. The vaccinia recombinant virus (V-RG) expresses the rabies glycoprotein. Both vaccines induce similar humoral and cell-mediated responses after primary and secondary oral administration. We observed a typical anamnestic response, although of a limited duration, after the booster vaccination. Therefore, our results suggested that two successive oral vaccination campaigns should not significantly improve the immunisation of foxes. Lymphocyte in vitro proliferative response to the SAD antigen highlighted the presence in blood of a T-cell specific memory 6 months after vaccination. The synthesis of several vulpine cytokines was detected in peripheral blood mononuclear cells (PBMC) stimulated by SAD antigen via reverse transcription polymerase chain amplification. The data showed a concomitant expression of interleukin (IL)-4 and interferon-gamma in PBMC of vaccinated foxes. No change was detected in the level of IL-2, IL-10 and IL-12 synthesis, whereas the pro-inflammatory cytokine tumour necrosis factor-alpha seemed involved in the activation of naive T lymphocytes.

Administration, Oral↗

A parasitological survey of wild red foxes (Vulpes vulpes) from the province of Guadalajara, Spain.

An epizootiological survey of leishmaniosis, coccidiosis and parasitic helminths in 67 foxes (Vulpes vulpes) was conducted in Guadalajara (central Spain). Examination for parasitic protozoa revealed prevalences of 74% Leishmania (determined by molecular methods) and 2.9% coccidia oocysts (fecal flotation). Survey of parasitic helminths (fecal flotation/necropsy) demonstrated the presence of nine species, including six nematodes, two cestodes and one trematode. Nematodes were the most common parasites of foxes, followed by cestodes and trematodes. Greater levels of nematodes like Uncinaria, with a free-living stage in its life-cycle, were found in foxes in areas where moist soils were likely to exist, in contrast to areas of semiarid characteristics, where Toxascaris leonina or Trichuris vulpis were predominant. With regard to helminths of importance as human pathogens, trichinoscopy revealed the presence of a relatively high number of foxes (8.9%) infected with Trichinella spiralis. Finally, Toxocara canis infection was less frequent (4.4%) than trichinellosis.

Animals↗

Seroprevalences of Toxoplasma gondii and Neospora caninum in Swedish red foxes (Vulpes vulpes).

The prevalences of antibodies to the protozoan parasites Toxoplasma gondii and Neospora caninum were investigated by the direct agglutination test (DAT) and ELISA, respectively, in 221 red foxes (Vulpes vulpes) from different parts of Sweden. A total of 84 (38%) of the analysed sera had antibodies to T. gondii, but none of the foxes had antibodies to N. caninum. The results indicate that T. gondii infection is fairly common in Swedish red foxes and that the infection is present in most parts of the country. They also show that N. caninum is not widespread as a latent infection among red foxes in Sweden.

Agglutination Tests↗

Ectoparasite infestations of red foxes (Vulpes vulpes) in Hungary.

A survey was carried out to investigate the ectoparasite infestations of 100 red foxes (Vulpes vulpes) in Hungary. The overall prevalence of flea and tick infestation was high (62 and 86% with the dominance of the anthropophil Pulex irritans and Ixodes ricinus), but the number of parasites was low to moderate. Felicola vulpis was not found in the present study, and the prevalence of Otodectes cynotis was only 2%. Based on prevalence (21%), mange lesion scores, and the negative correlation between lesion scores and condition of foxes, Sarcoptes infestation should be considered as the most important parasitosis of foxes in Hungary. Besides the ecological significance of these parasites, the high overall prevalence of mange and anthropophil flea and tick infestations of foxes and the appearance of these animals in the synanthropic environment as a result of the increasing population size, may result in the increasing incidence of flea, tick and accidental mite infestation of man and domestic animals, and may enhance the transmission rate of some vector-borne diseases.

Animals↗

Canine heartworm (Dirofilaria immitis) detected in red foxes (Vulpes vulpes) in urban Melbourne.

Canine heartworm (Dirofilaria immitis) was detected by antigen ELISA in 8 (6.4%) of 125 red foxes (Vulpes vulpes) captured in the Melbourne (Australia) metropolitan area. Circulating microfilariae were also detected by whole blood filtration in six of these foxes. Cursory sampling of mosquitos at two of the sites where positive foxes were captured revealed the existence of two known mosquito vectors of Dirofilaria. The existence of widespread fox populations in Melbourne, together with known mosquito vectors may offer the potential for a sylvatic cycle of canine heartworm within the urban area.

Aedes↗

First report of Trichinella nativa in red foxes (Vulpes vulpes schrencki) from Otaru City, Hokkaido, Japan.

Forty-three red foxes (Vulpes vulpes schrencki) and nine raccoon dogs (Nyctereutes procyonoides viverrinus) were captured in Otaru City, Hokkaido, Japan and examined by muscle digestion for the presence of Trichinella sp. larvae. Of the foxes, five (11.6%) were positive for larvae of Trichinella nativa while none of the raccoon dogs were found to be positive. This finding suggests that the red foxes are important reservoir hosts of sylvatic trichinellosis in Otaru, Hokkaido. This is the first report on Trichinella nativa in wild red foxes of Hokkaido, Japan.

Animals↗

Coproantigen survey for Echinococcus multilocularis prevalence of red foxes in Hokkaido, Japan.

An epidemiological survey was conducted on the seasonal variation of Echinococcus multilocularis prevalence in red foxes from 1997 to 1998, using a monoclonal antibody-based detection of the tapeworm coproantigen. Thirty-six breeding dens of reproductive fox families were identified in the endemic area of Koshimizu, eastern Hokkaido, Japan. Fecal samples from each site were examined by coproantigen detection assay and fecal egg examination. Whereas the prevalence of coproantigen positive feces showed no seasonal fluctuation (51.6-66.7%), variation was found in the prevalence of egg positive feces in which a higher prevalence was observed in the summer and winter (31.1 and 38.7%) than spring and autumn (13.3 and 13.5%). Significant differences were observed between juveniles and adult foxes in both examinations. Samples from juvenile foxes gave higher coproantigen positive results and taeniid egg intensity. Those results suggest more juveniles infected with the cestode than adults in the same period. The practical use of coproantigen assay as a survey tool and factors which affect the prevalence and host age-related difference are discussed.

Animals↗

Epidemiology of canine leishmaniasis: a comparative serological study of dogs and foxes in Amazon Brazil.

The paper describes a longitudinal field study of canine leishmaniasis in sympatric domestic dog and crab-eating fox (Cerdocyon thous) populations. Dogs were studied in house-to-house surveys, and foxes were studied by live-trapping and radio-telemetry. Because serological tests (IFAT in this case) for leishmaniasis are often of uncertain sensitivity and specificity, we draw conclusions comparatively. Both cross-sectional (age-prevalence) and longitudinal analyses indicate that incidence in dogs was highest in the dry season. Seasonal changes in the age-prevalence relationship for dogs suggest that serological conversion and recovery rates decline with prior exposure to infection, where 'recovery' may be due to loss of a positive antibody titre or death from leishmaniasis. The mean incidence in dogs was higher in the rural than in the urban population and higher in hunting dogs than pet dogs. Cross-sectional and longitudinal analyses of fox serological data suggest that foxes remain positive for longer than dogs on average, either because detectable antibody is more persistent or because they experience a lower mortality rate due to leishmaniasis.

Animals↗

Hammondia isolated from dogs and foxes are genetically distinct.

Hammondia heydorni is regarded as a protozoan parasite that uses canids, e.g. dogs and foxes, as definitive hosts, but clinical signs of infection are rare. This study therefore took advantage of the opportunity to study an oocyst population from the faeces of a dog suffering from intermittent bouts of diarrhoea. Oocysts from the naturally infected dog were shown to be H. heydorni by using the polymerase chain reaction combined with DNA sequencing as a diagnostic tool. The nucleotide sequence data reported in this paper are available from GenBank under the following Accession numbers DQ183058, DQ183059 and DQ022687. A comparison of the first internal transcribed spacer (ITS1) sequence of ribosomal DNA obtained with those from other dog and fox oocysts, previously regarded as H. heydorni, showed these oocysts contained identical ITS1 sequences. However, the oocyst DNA from the fox and dog differed by the presence/absence of a 9 bp insertion/deletion within intron 1 of the alpha tubulin gene, and this difference was conserved across a number of different oocyst populations from the 2 species of host. A PCR assay was established that takes advantage of this insertion/deletion and is able to differentiate between the 2 oocyst populations. This study therefore provides evidence that H. heydorni oocysts from dogs and foxes represent 2 distinct genetic lineages that can be differentiated using a PCR, which targets the alpha tubulin locus.

Animals↗

Polyploidization in the trophoblast and uterine glandular epithelium of the endotheliochorial placenta of silver fox (Vulpes fulvus Desm.), as revealed by the DNA content.

Dynamics of genome multiplication during establishment of interrelations between trophoblast and glandular epithelium of the endometrium has been studied in the course of formation of placenta in the silver fox. During formation of the placenta, penetration of the trophoblast into the zone of the endometrial glandular epithelium and of endometrial blood vessels into the zone of expanding trophoblast occurs. The trophoblast, which gradually replaces epithelium and a part of the stroma of the endometrium, closely adjoins endometrial vessels but does not disrupt them, thereby the endotheliochorial placenta is formed. Cytophotometric measurements of the DNA content in trophoblast nuclei have shown that most of them are polyploid: predominantly 4-64c, occasionally 128c and 256c. Polyploidy of the trophoblast may be a consequence of various types of polyploidizing mitoses. Cytophotometric measurements of the DNA content in mitotic figures have revealed the presence of mitoses of diploid cells, i.e. with the DNA amount of 4c (2n), and polyploid cells, i.e. 8c (4n), and 16c (8n), therefore trophoblast cells in the silver fox placenta are able to enter mitosis up to the octaploid level. Higher degrees of polyploidy in the trophoblast cells seem to be achieved by endoreduplication. Polyploidization of the uterine glandular epithelial cells during placentation in the silver fox occurs until the level of 8c. Thus, the tissue-specific response of the uterus to the implanting embryo consists of active proliferation and polyploidization of the glandular epithelium, which may compensate formation of prominent population of decidual cells (i.e., connective tissue cells). In the endotheliochorial placenta of the silver fox the regularity is confirmed that cells of both maternal and fetal origin are, as a rule, polyploid in sites of their contact in placenta, which may be of protective significance in the contact of allogenic organisms.

Animals↗

Can foxes be controlled by reducing their fertility?

A model based on data from research in New South Wales conducted by the Cooperative Research Centre for the Biological Control of Vertebrate Pest Populations suggests that the effectiveness of fertility control in reducing the abundance of foxes (Vulpes vulpes) can be strongly influenced by environmental variability. The model includes age-specific recruitment and survival as functions of resources indexed by rainfall. It is assumed that fertility control will affect only female foxes and that the use of a baiting regime to deliver a contraceptive agent will result in fixed proportional changes in pregnancy rates. By comparing the variability in the rate of increase of treated and untreated fox populations, the model predicts that: (i) frequent baiting, every one or two years, will be more effective than applications of baits at longer time intervals; (ii) the abundance of foxes will decline more rapidly, with higher levels of fertility control; (iii) infertility which lasts for only one breeding season is less effective than permanent sterility which allows for accumulation of sterile animals in the population; and (iv) highly variable results are likely to be the outcome of low-frequency baiting with an agent that produces only temporary infertility.

Animals↗

A bait-delivered immunocontraceptive vaccine for the European red fox (Vulpes vulpes) by the year 2002?

An orally-delivered immunocontraceptive vaccine is being developed for the control of fox populations. A number of genes (PH-20, LDH-C4, ZP3) encoding gamete proteins have been cloned, produced in recombinant expression systems and used in fertility trials to test the efficacy of these antigens. As the immunocontraceptive vaccine will be delivered in a bait, there is a requirement for a greater understanding of the immune responses of the reproductive mucosa in canids, and the assessment of the best vaccine delivery system that will evoke a mucosal antibody response. Several vaccine delivery systems including microencapsulated antigens, and both vaccinia virus and bacterial vectors are being investigated. Oral administration of Salmonella typhimurium recombinants expressing different fox sperm antigens stimulates both systemic IgG responses to the antigen and a mucosal immune response within the female reproductive tract in the fox, indicating that salmonella may have potential with respect to the oral delivery of antigen. The enhancement of mucosal immune responses to orally-delivered vaccines is also being examined, research focussing on the possible use of fox-specific cytokines or the beta-subunit of cholera toxin in forming part of the vaccine construct.

Adjuvants, Immunologic↗

Construction and immunological assessment of Salmonella typhimurium expressing fox sperm LDH-C4.

This study examined immune responses of foxes to oral doses of recombinant Salmonella typhimurium expressing fox sperm-specific lactate dehydrogenase (fLDH). The cDNA for fLDH was cloned into the expression plasmid pKK233.2 (pKKfLDH). Salmonella typhimurium aroA- (SL3261) was transformed with either the pKK233.2 plasmid alone (SpKK) or the pKKfLDH construct (SpKfLDH). The fLDH expressed by SpKfLDH retained enzymatic activity and was recognized by human LDH-C4-specific antibody. Male European red foxes (Vulpes vulpes) were given an initial oral dose of 1 x 10(11) cfu of either SpKK (control, n = 3) or SpKfLDH (test, n = 6), followed four weeks later with a further dose of 1 x 10(11) cfu. Antibodies to Salmonella lipopolysaccharide (LPS-04) and fLDH were measured in plasma and saliva for eight consecutive weeks after the initial doses. Both LPS-04 IgG- and IgA-specific antibodies as well as fLDH-specific IgG antibodies were detected in plasma and saliva. However, there was a marked fLDH-specific IgA response in saliva consistent with induction of the common mucosal immune system. The antibody measurements demonstrated the feasibility of using recombinant Salmonella as an oral vaccine to elicit gamete antigen-specific mucosal immune responses in foxes.

Amino Acid Sequence↗

Cloning and characterization of the cDNA encoding the PH20 protein in the European red fox Vulpes vulpes.

The PH20 protein is thought to play a crucial role in mammalian fertilization. The fox PH20 homologue has been cloned from a testis cDNA library and the deduced protein sequence shows high levels of homology to PH20 proteins isolated from other species. Unlike other PH20 proteins the fox protein does not appear to be membrane associated through a GPI-linkage nor does it show the presence of a transmembrane domain at the C-terminus of the protein. It is in this region that the proteins appear to be least conserved. Immunolocalization studies on fox sperm show that the PH20 protein is located on the inner acrosomal membrane. Transcription of PH20 in the fox is seasonally regulated, with the mRNA expressed during those months when spermatogenesis is at its peak. The PH20 sequence described in this paper has been submitted to the Genbank database and has the accession number U41412.

Amino Acid Sequence↗

Alterations of the pituitary-gonadal axis in the neonatal blue fox (Alopex lagopus) exposed to prenatal handling stress.

Handling is a stressor for blue foxes. The influence of preterm handling (1 min daily) of vixens was investigated in 68, 10-day-old cubs, 34 male and 34 female. Body, gonadal and pituitary weight, ano-genital distance, gonadal testosterone and oestradiol content and in vitro production, and pituitary LH content were measured in all cubs. The gonads were frozen or incubated in vitro with, or without, added hCG (2.5 IU per sample). The gonadal incubates and homogenates were analysed for testosterone and oestradiol by radioimmunoassay and the pituitary homogenates for LH by immunofluorometric assay. The results indicate that neonatal fox gonads actively produce steroids and that there are significant sex differences in basal steroid production and response of the gonads to hCG, and in pituitary LH content. Maternal stress resulted in a significant reduction of morphometric and hormonal measures of the reproductive system in neonatal blue foxes, with more drastic effects in female cubs. Gonadal weights were lower in cubs of both sexes from stressed vixens (65.7+/-4.3 v 50.6+/-1.8 mg for the ovaries and 23.2+/-1.0 v. 17.7+/-1.0 mg for the testes, control v. stressed animals, P<0.01). The ano-genital distance in female offspring of stressed vixens was reduced (1.1+/-0.04 v. 0.9+/-0.03 cm, P<0.01). Basal ovarian oestradiol and testosterone production were decreased in cubs from prenatally stressed animals in comparison with controls (43.5+/-3.5 v. 32.6+/-3.7 pg ovary(-1) h(-1) and 0.40+/-0.16 v. 0.12+/-0.03 ng ovary(-1) h , P<0.05). Prenatal stress did not affect either pituitary weights or LH content in either sex. There were no significant differences in ano-genital distance, testicular content of testosterone, or in vitro testosterone production between control and treated male cubs. In conclusion, these findings suggest that prenatal handling stress impaired the neonatal reproductive development of the female offspring, but had no marked effects on males. Sex-specific effects of prenatal handling stress on the reproductive development in foxes may be linked with the gender differences in responses of the hypothalamus-pituitary-adrenocortical axis to stress conditions in this species.

Adrenal Cortex↗

Cloning and characterization of a fox sperm protein FSA-1.

A monoclonal antibody was raised to a fox sperm protein (FSA-1) which was found to be localized to the inner acrosomal compartment of sperm fixed in methanol. Western blots of testicular germ cell membrane extracts probed with this antibody identified a major protein band with a molecular weight of 36,000. Immunofluorescent studies on fox testis sections showed that the antigen is expressed on round and elongating spermatids on a crescent-shaped structure, which probably represents the developing acrosome. An antibody specific for FSA-1 was used to screen a fox testis cDNA library for its cognate gene. An 875-bp cDNA clone was isolated and sequenced revealing an open reading frame. Searches of the GenBank and EMBL databases with the nucleic acid sequence revealed significant homology (86%) of FSA-1 with 406 bases of an unidentified RNA transcript from human fetal brain (EST02625). Northern blot analysis of fox testis RNA samples identified an RNA transcript of approximately 0.9 kb during the months when spermatogenesis is active. Zoo Northern blots (at high stringency) reveal an RNA transcript of a similar size present in testis RNA from dogs and mice. Zoo Southern analysis (high stringency) reveal genomic sequences present in dogs, mice, cattle and sheep. At present, the function of the FSA-1 gene product remains unknown, but it may play a role as a structural protein component of the acrosome.

Acrosome↗

The growth and development of Echinococcus granulosus of sheep origin in dogs and foxes in Britain.

Four separate experiments were carried out by dosing dogs (Beagles and Border collies) and red foxes with protoscoleces obtained from hydatid cysts in sheep from mid-Wales. The dogs and foxes were killed at intervals over a period from 34-180 days after infection and the intestines were examined for Echinococcus granulosus. Border collie dogs were efficient hosts in that large numbers of worms developed, with gravid segments present by 55 days after infection; but Beagle dogs were poor hosts in that only small numbers of immature worms developed. Foxes were poor hosts in that few or no worms developed, but gravid segments were seen in three of 29 foxes. The worms showed morphological features similar to the 'sheep' rather than the 'horse' strain.

Animals↗

Chemotherapy with praziquantel has the potential to reduce the prevalence of Echinococcus multilocularis in wild foxes (Vulpes vulpes).

In order to evaluate the applicability of anthelminthic treatment of wild foxes (Vulpes vulpes) to limit their infection with Echinococcus multilocularis, bait pellets, each containing 50 mg praziquantel, were repeatedly distributed in an area of 566 km2 where many foxes are infected, in southern Germany. After six baiting campaigns (15-20 baits/km2) over a period of 14 months, the prevalence of the cestode in foxes, initially 32%, had fallen to 4%. The effect was most pronounced in the central part of the treated area, where no positive fox was found in the 2 months before the end of the trial. The study was controlled for other factors that could influence the parasite's prevalence, such as the availability of intermediate hosts. While the potential of this baiting method to remove E. multilocularis from wild hosts has been demonstrated, the question of its long-term efficacy and other unresolved problems have to be addressed by consecutive studies before routine application can be recommended.

Animals↗