An indirect ELISA for detection of Encephalitozoon cuniculi infection in farmed blue foxes (Alopex lagopus)

Infection with the intracellular microsporidium Encephalitozoon cuniculi can cause serious disease, encephalitozoonosis, in the blue fox (Alopex lagopus). The disease diagnosis is based on clinical signs and pathological findings, and detection of E. cuniculi or circulating antibodies directed against the parasite. Indirect immunofluorescence (IFAT) and carbon immunoassay (CIA) are the most commonly used serological methods for diagnosis in this species. In the present study, an indirect ELISA (enzyme linked immunosorbent assay) was established and evaluated against IFAT by testing of 205 field samples from blue foxes. There was high agreement between the results of the ELISA and CIA (kappa=0.99), and the ELISA and IFAT (kappa=0.958). There was no significant statistical difference between the tests (p>0.05). It was concluded that the ELISA could be used to identify seropositive farmed blue foxes. The advantage of the ELISA lies in the potential of screening large numbers of animals with the goal of eradicating E. cuniculi infection in the farms.     

Medetomidine- and medetomidine-ketamine-induced immobilization in blue foxes (Alopex lagopus) and its reversal by atipamezole

The sedative and immobilizing effects of the alpha 2-adrenoceptor agonist medetomidine alone or combined with the dissociative anesthetic ketamine, were studied in blue foxes. Medetomidine at doses of 25 and 50 micrograms/kg induced moderate to deep sedation, but only with the highest medetomidine dose tested, 100 micrograms/kg, was the immobilization complete. Medetomidine 50 micrograms/kg combined with ketamine 2.5 mg/kg rapidly induced complete immobilization, characterized by good myorelaxation, and no clinically significant alterations in serially determined hematologic and serum chemistry parameters. The alpha 2-adrenoceptor antagonist atipamezole effectively reversed the medetomidine- or medetomidine-ketamine-induced immobilizations. A transient increase in heart rates was noted after each atipamezole injection.     

Cholinesterase activities in uterus of normal and fenchlorphos treated blue foxes (Alopex lagopus) during various reproductive states

The uterine acetylcholinesterase and total cholinesterase (acetylcholinesterase plus butyrylcholinesterase) activities in normal and fenchlorphos treated blue fox vixens were determined during various reproductive states.AChE and Total-ChE of non-medicated vixens in oestrus were about one half of those in anoestrus. In pregnant uteri (luteal phase) the activities were 25 % and 30% compared to anoestrus.In vixens given 100 mg/kg fenchlorphos for 3 weeks during anoestrus, the remaining activity of AChE in uterus were in average 37%. Pregnant and non-pregnant vixens in the luteal phase medicated prior to mating and during time of implantation, displayed AChE activities which were only moderabely reduced (remaining activities 83% and 72% compared to medicated animals in anoestrus: remaining activity 37%).Plasma ChE-activity increased during pregnancy in the controls while enzyme activity was strongly reduced in animals given 100 mg/kg fenchlorphos daily through the whole pregnancy.It was concluded that the previous reported embryotoxic effect of fenchlorphos in the blue fox did not seem to be directed towards the moderate inhibition of the uterine cholinesterases.     

Humoral immune response in adult blue foxes (Alopex lagopus) after oral infection with Encephalitozoon cuniculi spores

Encephalitozoon cuniculi causes severe diseases in blue fox puppies. When pregnant vixens are infected, parasites are transmitted over the placenta to the unborn that subsequently develop encephalitozoonosis. Adult foxes themselves do not have signs of disease, but show antibody titres to E. cuniculi. The purpose of the present study was to gain information on the immune response in adult foxes after experimental infection. Sixteen foxes were infected orally with E. cuniculi spores, eight of them twice and 28 days apart. The two groups of animals showed elevated serological values in both the carbon immunoassay and in the ELISA. Elevated serological levels were recorded up to 1 year after the infection took place. The control group (n=8) remained serologically negative throughout the trial. The results of the study showed that blue foxes could be seropositive for at least a year after oral infection with E. cuniculi.     

Effects of dietary fat levels on nutrient digestibility and production performance of growing-furring blue foxes (Alopex lagopus)

The objective of this study was to determine whether nutrient digestibility and production performance of growing-furring blue foxes (Alopex lagopus) are affected by different dietary fat levels. Sixty-four young animals were randomly assigned to four groups (A, B, C, D) provided with diets containing approximately 12%, 26%, 40%, 54% fat in the dry matter respectively. When dietary fat level was increased, the apparent digestibility of main nutrients except for crude carbohydrates, and gross energy were improved (p < 0.0001). The amount of nitrogen excreted was reduced and the biological value of protein was enhanced (p < 0.0001). Over the experimental phase, the efficiency of metabolizable energy (ME) used for gain in group B (26% fat content) was higher than that in other groups. When diets with 12-26% fat content were fed, there was an increasing tendency in skin size. But it had negative effects on skin size and fur quality when the amount of fat content was over 40%. In conclusion, the experiment showed that dietary fat could significantly improve some nutrient utilization and significantly reduce feed/gain ratio as a main energy source. The most preferable fur quality and efficiency of ME used for gain were obtained when diet contained 26% fat level in growing-furring period.     

An outbreak of septic endometritis in the Arctic blue fox (Alopex lagopus) caused by Clostridium carnis

An outbreak of endometritis and septicaemia in farms that raise Arctic blue foxes is described. Eleven animals from eight different farms were examined, most of them pregnant or with newly delivered litters. Pathological examination indicated inflammation of the uterus, and focal necrotic changes in the liver. In some animals evidence of hemorrhaging was found in the anterior part of the small intestine. Bacteriological examination revealed abundant growth from different organs of the animals of a pure culture of a small, slender, Gram-variable, sporeforming rod, which was identified phenotypically as aerotolerant Clostridium carnis. This identification was confirmed by DNA hybridization. The organism caused death in mice 1–2 days after intraperitoneal inoculation, and killed an Arctic blue fox which was inoculated intracervically. Two foxes, pretreated with diethyl-stilboestrol, became seriously ill after intrauterine inoculation, but survived. The spontaneous outbreak of the disease was characterized by sudden onset of septicaemia with few or no clinical symptoms before the animals succumbed.     

Detection of herpesvirus DNA in Arctic foxes (Vulpes lagopus; syn. Alopex lagopus) with fatal encephalitis

A captive breeding programme for the Fennoscandian Arctic fox (Vulpes lagopus; syn. Alopex lagopus) failed due to fatal encephalitis. The aim of this study was to identify the causative agent. Viral nucleic acid was detected by PCR and in situ hybridization in the brain of affected foxes. The results suggest that a herpesvirus might be the causative agent. Whether this infection also occurs in free-living Arctic foxes is unknown.     

蓝狐MITF-M基因序列扩增及生物信息学分析

试验旨在研究蓝狐MITF-M基因核心启动子活性区、转录因子结合位点、基因编码蛋白结构及功能,为探究该基因的表达调控机制提供理论依据。从成年蓝狐背部皮肤组织中采集1 cm×1 cm样品,采用RT-PCR扩增及测序技术首次获得MITF-M基因序列3 880 bp（包括5'侧翼区2 262 bp、CDS区1 260 bp、3'侧翼区358 bp）,编码419个氨基酸残基。生物信息学分析结果显示,蓝狐MITF-M基因5'侧翼区存在潜在的启动子区域（-86~-336 bp）,且发现TATA框、CAAT框、GC框和CRE等调控元件及CREB、LSF和Sp1等多种转录因子。MITF-M基因编码的蛋白是定位于细胞核和细胞质的不稳定、可溶性亲水蛋白质。亚细胞定位结果提示该蛋白在细胞核、细胞质和线粒体的概率较高,分别为65.2%、17.4%和8.7%。DNAMAN软件对MITF-M基因编码蛋白的二级结构预测发现,该蛋白由α螺旋（33.66%）、β折叠（16.66%）和无规卷曲（49.68%）组成,且不存在跨膜区域和信号肽。蓝狐与其他物种MITF-M基因编码的氨基酸序列同源性对比和系统进化树分析均提示蓝狐与犬的遗传亲缘关系最近,与哺乳动物和禽类均具有较高的同源性（>89.1%）,说明MITF-M基因编码区在物种进化过程中较为保守。本研究为深入研究MITF-M基因调控狐狸被毛颜色的分子遗传机制奠定了理论基础。     

北极狐酪氨酸酶基因cDNA序列分析及启动子预测

为了探明北极狐酪氨酸酶(tyrosine,TYR)基因核心启动子活性区、转录因子结合位点、基因编码蛋白结构及功能,通过RT-PCR扩增及基因测序技术首次从北极狐皮肤组织中获得TYR基因c DNA序列,全长4 300 bp(包括5'侧翼区2 681 bp,CDS区1 593 bp,3'侧翼区26 bp)。生物信息学分析结果表明:北极狐TYR基因5'侧翼区存在潜在的启动子区域,并发现TATA框和CAAT框等调控元件及髓细胞增生原癌基因(Myc)、胰十二指肠同源盒1(Pdx1)、V-Myb髓细胞组织增生病毒癌基因同源物(Myb)、E26转录因子1(Ets1)、特异性蛋白1(Sp1)和TATA盒结合蛋白(TBP)等多种转录因子。北极狐TYR基因编码蛋白的长度为530个氨基酸残基,编码蛋白是定位于生物膜上的不稳定可溶性亲水蛋白质,该蛋白以无规卷曲为主,主要参与信号转导和转录,存在跨膜区域和信号肽。北极狐与其他物种的系统进化树分析提示北极狐与家犬的遗传亲缘关系最近,不同物种间亲缘关系与生物进化观点基本一致,符合动物学分类。     

北极狐电刺激采精技术的研究

1995～ 1 997年北极狐配种期对 41只雄狐进行 97次电刺激采精试验 ,成功 96次 ,成功率 98 9%。雄狐用安定注射液和盐酸氯胺酮进行复合麻醉。电刺激采精器频率调至 2 0～ 30Hz;通、断电间隔时间 7s;电压由 2V开始 ,进行电刺激 ,每持续 30s后再升高 2V ,直至升到 30～ 32V公狐即可排精。所得到精液品质优良 ,采精后公狐食欲、精神均正常。     

Surgical implantation of radio transmitters in arctic foxes (Alopex lagopus) on Svalbard, Norway.

Twelve captive wild-caught adult arctic foxes (Alopex lagopus) were anesthetized a total of 24 times for an equal number of surgical procedures involving implantation of heart rate (HR) and core body temperature transmitters (Tb) between October 1995 and April 1997. Xylazine-ketamine and medetomidine-ketamine anesthesia was used, resulting in an unacceptably high death rate. One out of four foxes anesthetized with xylazine-ketamine died, whereas two of nine foxes anesthetized with medetomidine-ketamine died out of a total of 20 surgical procedures. Durations of the surgeries for implantation of Tb transmitters and HR transmitters were 73 +/- 7 min and 95 +/- 13 min, respectively.     

Periovulatory endocrinology and oocyte maturation in unmated mature blue fox vixens (Alopex lagopus)

Nine of 10 mature blue fox vixens (Alopex lagopus) in spontaneous oestrus ovulated approximately 2 days after the preovulatory increase in luteinizing hormone (LH). Plasma concentrations of follicle-stimulating hormone and progesterone increased simultaneously with the LH peak, whereas oestradiol-17 beta peaked 1 day previously. In the tenth vixen, an LH peak was not observed, and neither visible follicles nor corpora lutea were found in the ovaries 6 days after peak vaginal electrical resistance. Eggs were ovulated as primary oocytes, but oocyte maturation was initiated within the day of ovulation (2 days after the LH peak). Within the next 2 days (3-4 days after the LH peak) the first polar body was extruded, and the cumulus mass was completely dissociated from the zona pellucida. The interval between the preovulatory LH peak and initiation of the final oocyte maturation is thus considerably longer in the blue fox than for example in the cow (48-72 h compared with 9-12 h). This suggests that the relationship between these two events is somewhat different in the blue fox.     

Neuropathology and host-parasite relationship of acute experimental toxoplasmosis of the blue fox (Alopex lagopus)

The neuropathology and host-parasite relationship of experimental infection with the RH-strain of Toxoplasma gondii were studied in 27 blue foxes (Alopex lagopus) aged 0 to 23 days, using light microscopy, including immunohistochemical staining, and transmission and scanning electron microscopy. All cases displayed multifocal necrotic lesions with numerous parasitic tachyzoites in the brain and spinal cord. The gray matter and the meninges were most seriously affected. Although a wide variety of cell types were parasitized, neurons and astrocytes seemed to be the main target cells. Individual parasitophorous vacuoles usually contained only a few tachyzoites, with rosette formations as a prominent feature. The present ultrastructural study supports the theory that the parasites actively invade the host cells by mechanisms that are different from those of phagocytosis. This is apparently the first report indicating that the formation of the network of tubular structures within the parasitophorous vacuole of T. gondii is associated with a transient, sack-like formation in the posterior end of the tachyzoites.     

育成期舍饲蓝狐结肠菌群多样性分析

旨在分析蓝狐结肠菌群的组成及其多样性,为蓝狐肠道菌群体外培养提供基础数据。随机选取6只健康育成期(3～4月龄)雌性舍饲蓝狐,采集结肠食糜后采用高通量测序技术对蓝狐结肠菌群结构进行分析。结果表明:6个样品共获得377 798条有效序列,平均为62 966条有效序列;共获得2 064个OTU,平均为307个OTU,归类于12菌门、180个菌属。在门分类水平上,以厚壁菌门(Fir-micutes,70.25%)、拟杆菌门(Bacteroidetes,22.36%)、变形菌门(Proteobacteria,5.69%)、放线菌门(Acti-nobacteria,1.13%)为主,4个菌门占所有细菌总数的99.43%。在属分类水平上,以乳杆菌属(Lactoba-cillus,28.50%)、巨单胞菌属(Megamonas,7.48%)、拟普雷沃菌属(Alloprevotella,7.41%)、Peptoclostridi-um(6.33%)、乳球菌属(Lactococcus,5.38%)、Dialiste (4.39%)、巨型球菌属(Megasphaera,3.53%)、De-sulfovibrio(2.97%)、拟杆菌属(Bacteroides,2.48%)、unidentified_Lachnospiraceae(2.38%)为主。结果表明,在饲养条件下,蓝狐结肠中具有相对复杂的菌群结构,且个体间存在较大差异,研究为课题组进一步揭示蓝狐结肠菌群功能以及探索蓝狐结肠菌群体外培养研究提供了基础数据。     

Scanning Electron Microscopic Study of the Lingual Papillae in the Arctic Fox (Alopex lagopus L., 1758)

This study aims to show the distribution and the three-dimensional structure of the lingual papillae in the arctic fox. The macro- and microscopic structure of the tongue and its lingual papillae was studied in 11 adult arctic foxes. Two types of mechanical papillae were distinguished on the dorsal surface of the tongue – filiform papillae and conical papillae. The gustatory papillae in the arctic fox are represented by fungiform, vallate and foliate papillae. The keratinized filiform papillae on the anterior part of tongue are composed of one big posterior process accompanied by 10–12 secondary anterior processes. The number of anterior processes of filiform papillae undergo a complete reduction within the area between the posterior part of the body of the tongue and area of the vallate papillae. The conical papillae cover the whole dorsal surface of the root of the tongue, including the lateral parts surrounding the area of the vallate papillae and the posterior part of the root. The size of the conical papillae increases towards the root of the tongue but their density decreases. In the arctic fox, there are three pairs of vallate papillae distributed on the plan of a triangle. The diameter of vallate papillae in each successive pair is bigger. The wall surrounding the body of the vallate papilla and its gustatory trench is composed of six to eight conical papillae joined at various degree. The foliate papillae on both margins of the tongue consist of seven to nine laminae.     

Detection of estrus and determination of the optimal time for insemination in polar foxes (Alopex lagopus)]

Oestrus cycles were investigated in seven polar foxes: the investigation consisted of evaluation of clinical symptoms of heat (vulva enlargement), measurement of vaginal mucus electrical resistance and determination of progesterone concentrations in the blood plasma of foxes; the objective was to determine optimum insemination time. The females were inseminated with fresh semen intravaginally by means of a pseudopenis, a day following the record of maximum electrical resistance of vaginal mucus and at the minimum progesterone concentration in blood plasma of 30 ng.ml-1. The females were reinseminated on the following day. The insemination dose had a volume of 1 ml with sperm concentrations of 150.10(6). Clinical symptoms of heat were observed in six out of the seven test foxes. Heat detection by means of measuring vaginal mucus electrical resistance was successful also in six females. Progesterone test enabled to detect heat in five females. Out of six inseminated females, five foxes became pregnant (83.3%) and a total of 33 cubs were born; this is 5.5 cubs per female.     

Ovarian and testicular function in the blue fox (Alopex lagopus) after oral administration of fenchlorphos during the breeding season

The possible effect of fenchlorphos, 0-0-dimethy1-0-(2.4.5-trichlorophenyl) phosphorothioate, upon the reproductive endocrinology in blue foxes (Alopex lagopus) was investigated. Five females were administered fenchlorphos orally at a dose of 100 mg/kg daily from 10 days before oestrus and up to the 21st day of gestation. This dose represents the therapeutic dose for the treatment of sarcoptic mange. Blood samples were collected for the analyses of progesterone, oestradiol-17β and luteinizing hormone (LH) in plasma. The vixens were ovario-hysterectomized on day 23, except 1 animal in the control group which was operated on day 17. Additionally, sperm quality and mating performance in 3 male blue foxes, which were administered 100 mg/kg fenchlorphos daily during the first 3 weeks of the mating season, were examined.Pregnancy was recorded in 2 medicated and 4 control animals. No pathological changes were observed in the uterus and the ovaries. The plasma concentrations of the hormones were similar to those obtained from the control group. No evidence of any disturbances concerning spermatogenesis in the males was observed. However, their libido appeared to be reduced. None of the males achieved a mating during and after the period of medication.     

Comparison between microscopic and automated differential leukocyte counts in the silver fox (Vulpes vulpes) and the blue fox (Alopex lagopus)

Differential leukocyte (WBC) counts in blood from clinically healthy silver foxes (n=32) and blue foxes (n=37) obtained from an automated hematology analyzer (Technicon H*1 Hematology System) with canine software were compared with microscopic differential WBC counts (M-diff). There was good agreement between the automated differential cell count (A-diff) and the M-diff for neutrophil and lymphocyte percentages. The correlation was lower for monocyte percentages and variable for eosinophil percentages. There was no significant difference between the A-diff and M-diff in either fox species. The A-diff counts were very precise, and may be a good alternative to the traditional M-diff for screening populations of clinically healthy foxes or for studies on stress and animal welfare. Intercept values, however, indicated a constant bias that must be taken into account before interpreting results based on different methods of analysis