Serologic and molecular survey of horses to Coxiella burnetii in East of Iran a highly endemic area

There are few reports about Q fever in horse populations worldwide. This study aimed to detect the C. burnetii infection by serologic and molecular confirmation using commercial ELISA kit and real-time PCR in the East of Iran a region highly endemic. A total of 177 blood samples and 115 vaginal swabs were randomly collected from horses in East of Iran. The sera samples were analyzed for anti C.burnetii Ig G antibodies by a commercial ELISA kit and nucleic acid extraxted from vaginal samples were used to determine the C. burnetii DNA by real-time PCR assay. Antibodies were detected in 5.64 % (10/177) of sera samples and C. burnetii DNA was detected in 7.82 % (9/115) of horse vaginal samples. There was no significant difference in seroprevalence in different sex, age and breed groups. Our study showed that horses could be considered as a mild potential reservoir of C. burnetii which may be effective on horse health status. However, additional studies are needed to assess whether the horse could be considered as a relevant transmission risk indicator for Q fever.     

菜豆萎蔫病菌的血清检测鉴定技术研究

本研究对我国一类检疫性有害生物菜豆萎蔫病菌（Ｃｕｒｔｏｂａｃｔｅｒｉｕｍｆｌａｃｕｍｆａ－ｃｉｅｎｓｐｖ．ｆｌａｃｕｍｆａｃｉｅｎｓ，Ｃｆ）进行血清检测鉴定技术的研究。试验中制备了抗Ｃｆ的血清，应用抗００２０，０２３７，０２３８菌株的抗血清及其ＦＩＴＣ标记的抗体进行间接和直接免疫荧光染色试验（ＩＦ／ＩＦ）。通过对１９个Ｃｆ菌株和其它６个属１４个种的植物病原细菌２０个菌株的测定，结果发现约５０％的Ｃｆ菌株为强阳性反应，且与Ｃｆ种下致病变种也有强阳性交叉反应。由于单个血清检测易产生假阴性反应，故将三种血清１∶１∶１混合后进行ＩＦ试验，结果表明与所有的Ｃｆ菌株反应，且与Ｒｆａ和Ｃｍｔ有弱阳性反应。对含１０４～１０７ｃｆｕ／ｍｌ细菌及３０ｇ种子含１粒人工接种种子的种子抽提液均能检测到典型的阳性细胞，血清检测的灵敏度为４．５×１０４ｃｆｕ／ｍｌ，可用于种子的检测。本研究还进行了初步的免疫荧光菌落染色（ＩＦＣ）试验     

Seroprevalence and Risk Factors Associated With Corynebacterium pseudotuberculosis Detectable Antibodies in Equids in Alabama

A cross-sectional serological survey was carried out to screen the equine population of the nonendemic state of Alabama for the presence of detectable antibody titers against Corynebacterium pseudotuberculosis. A second objective was to determine the association of detectable titers with risk factors such as exposure to ruminants or previous travel to endemic states. A total of 342 equine serum samples from 40 Alabama counties were analyzed using the synergistic hemolysis inhibition test (SHI). The prevalence of detectable antibody titers (≥1:8) was 52.5% (95% confidence interval [CI], 47%–57.9%). Titers ≥1:128 were detected in 2.63% (95% CI, 1.2%–4.9%), and titers ≥1:512 were detected in 0.3% (95% CI, 0%–1.6%) of the sampled population. In the final generalized linear model, age (P < .001), breed (P = .023), and contact with cattle (P = .05) were associated with increasing SHI titers. Contact with goats was associated in the initial but not in the final analysis (P = .19). Previous travel was not associated with increasing SHI titer (P = .97). The results demonstrated a high prevalence of low detectable titers and low prevalence of titers ≥ 1:128 in a nonendemic population. Further evaluation of SHI cutoff titers and accuracy is warranted to reduce the risk of a false positive diagnosis.     

First serological evidence of West Nile virus infection in wild birds in Northern Algeria

While the epidemiology of Flaviviruses has been extensively studied in most of the Mediterranean basin, little is known about the current situation in Algeria. In order to detect the circulation of West Nile (WNV) and Usutu viruses (USUV) in Kabylia, 165 sera were collected from two wild birds species, namely the long distance migrant Turdus philomelos (song thrush) (n = 92) and the resident Passer domesticus (house sparrow) (n = 73). A total of 154 sera were first analyzed by commercial competition ELISA. WNV and USUV micro-neutralization tests were performed on all c-ELISA positive sera and all samples with poor volume. Overall, 7.8 % (CI95 %: 3.5–11.9) were positive by c-ELISA. Positive results were detected in 12.5 % (CI95 %:5.6–19.4) of song thrushes and 1.5 % (CI95 %: 0.0–4.5) for sparrow.Micro-neutralization tests revealed an overall seroprevalence of 6.7 % for WNV (CI95 %: 2.9–10.3), Neutralizing antibodies were found in 8.7 % (CI95 %: 3.0–14.4) for song thrushes and in 4.1 % (CI95 %: 0.0–8.7) of sparrows. The current study demonstrates significant seroprevalence of WNV antibodies in wild birds in Algeria.     

Development and evaluation of an enzyme-linked immunosorbent assay for the serodiagnosis of pythiosis in dogs

Pythiosis (caused by the aquatic oomycete Pythium insidiosum) is a devastating and often fatal cause of either severe transmural gastroenteritis or locally invasive subcutaneous disease in dogs living in the southeastern United States. Although early diagnosis is essential for successful treatment, tools available for this task are limited. Therefore, we developed and evaluated an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-P insidiosum antibodies in canine serum. A soluble mycelial extract of P insidiosum was utilized as antigen in the ELISA, which was used to evaluate serum from 43 dogs with pythiosis, 8 dogs with lagenidiosis (another canine oomycosis), 16 dogs with nonoomycotic fungal or algal infections, 22 dogs with nonfungal gastrointestinal or skin disease, and 55 healthy dogs. Results were expressed as percent positivity (PP) relative to a strong positive control serum run on each plate. Medians and ranges for each of the 5 groups were as follows: pythiosis (81.7%, 50.6-98.5%), lagenidiosis (17.3%, 11.3-29.2%), other fungal or algal infections (8.2%, 4.7-15.4%), nonfungal gastrointestinal or skin disease (6.2%, 3.9-20.7%), and healthy dogs (6.7%, 3.0-15.2%). When using a cutoff value of 40% PP, the sensitivity and specificity of the ELISA both were 100%. In addition, ELISA values measured after successful surgical therapy in 2 dogs showed a decrease of anti-P insidiosum antibody concentrations into the normal range as early as 2 months after treatment. We conclude that the ELISA is a sensitive and specific test for the diagnosis of canine pythiosis, and may be a useful tool for monitoring response to medical or surgical therapy.     

A new furovirus infecting barley in France closely related to the Japanese soil-borne wheat mosaic virus

In April 2001, stunted barley plants bearing mosaic symptoms were observed in a field in France (Marne Department, 51). Rod-shaped and flexuous particles were visualized by electron microscopy and positive serological reactions were detected by ELISA with Barley yellow mosaic virus (BaYMV) and Soil-borne cereal mosaic virus (SBCMV) polyclonal antisera. The tubular virus which was soil transmissible to barley cv. Esterel was separated from BaYMV by serial mechanical inoculations to barley cv. Esterel. This furo-like virus, in contrast to a French isolate of SBCMV, could be transmitted to Hordeum vulgare, Avena sativa, Beta vulgaris and Datura stramonium. RT-PCR was used to amplify the 3′-terminal 1500 nucleotides of RNA1 and the almost complete sequence of RNA2. Nucleotide and amino acid sequence analyses revealed that the French virus infecting barley is closely related to a Japanese isolate of Soil-borne wheat mosaic virus (SBWMV-JT) which was originally isolated from barley. This French isolate was named SBWMV-Mar. The 3′ UTRs of both RNAs can be folded into tRNA-like structures which are preceded by a predicted upstream pseudoknot domain with seven and four pseudoknots for RNA1 and RNA2, respectively. The four pseudoknots strongly conserved in RNAs 1 and 2 of SBWMV-Mar show strong similarities to those described earlier in SBWMV RNA2 and were also found in the 3′ UTR of Oat golden stripe virus RNAs 1 and 2 and Chinese wheat mosaic virus RNA2. Sequence analyses revealed that the RNAs 2 of SBWMV-Mar and -JT are likely to be the product of a recombination event between the 3′ UTRs of the RNAs 2 of SBWMV and SBCMV. This is the first report of the occurrence of an isolate closely related to SBWMV-JT outside of Japan.     

桑萎缩病病原类菌原体的提纯、致病性及抗血清制备研究

用0．3mol／L甘氨酸缓冲液从病桑嫩叶提取病原类菌原体（MLO）,提取液上清经硅藻土柱层析分离,其洗脱液用高速离心浓缩,制备成部分纯化的病原MLO提纯物。采用显微注射技术,将提纯病原注射到介体昆虫体内,使病原在昆虫体内繁殖,再将获毒虫放饲在健桑苗上传毒接种,获得表现典型萎缩病症状的病株。病柔韧皮部筛管细胞的超薄切片在电镜下观察到大量病原MLO粒子,证实提纯MLO具致病性。同时用提纯病原为免疫抗原制备兔抗血清,抗血清用健桑提取液吸收后,获得仅与病桑抗原产生反应的特异抗血清,免疫电泳测验、仅产生一条沉淀带。     

Epidemiological Analysis of Leptospira spp. Infection in Equids from the Brejo Paraibano Microregion of Brazil

Leptospirosis is a zoonotic disease that occurs in humans and several animal species, including cattle, pigs, and horses, and it has a worldwide distribution. In equids, leptospirosis manifests through recurrent uveitis and abortion and other reproductive disorders. The objective of the present study was to investigate the epidemiological situation of Leptospira spp. infection in equids from the Brejo Paraibano microregion in the northeast of Brazil, that is, to assess the prevalence of and identify the major risk factors for infection. For this purpose, blood samples were taken from 257 equids (204 horses, 46 mules, and 7 donkeys), and a microscopic agglutination test (MAT) was used as serological analysis, with 24 serovars of Leptospira spp. as antigens. Antibodies were observed in 16.2% of horses, 13.0% of mules, and 28.6% of donkeys. Animals between the ages of 2.5 and 11 years had a risk of infection 0.4 times lower than those younger than 2.5 years old (odds ratio, 0.4; 95% confidence interval, 0.18-0.90). Further studies are needed to identify potential wild reservoirs and the main routes of infections.     

Cystic echinococcosis in the Falkland Islands

From 1991 to 1993, an investigation into the epidemiology of cystic echinococcosis (CE) was carried out in the Falkland Islands to evaluate the progress of the hydatid eradication campaign. The prevalence of CE in sheep was assessed using abattoir and farm slaughter data, and the exposure of dogs to the parasite was estimated using immunological techniques. A total of 59 466 sheep was examined at slaughter for E. granulosus and T. hydatigena cysts and the entire dog population of the Falkland Islands (n = 908) was examined by ELISA for the presence of specific serum antibodies to E. granulosus (IgG, IgA and IgE). In addition, a subsample of dogs (n = 464) was tested for the presence of E. granulosus antigens in faeces (copro-antigens). The prevalence of CE in sheep increased significantly during the period of the study from 0.11% in 1991 to 0.47% in 1993. Nineteen (2.1%) of 908 dog sera tested were seropositive, and eight dogs (1.7%) of 464 tested were positive in the copro-antigen assay. The combined use of abattoir surveillance, specific antibody and copro-antigen assay suggested that there were several locations in the Falkland Islands where the life cycle of E. granulosus may still perpetuate. Specific deficiencies in the eradication effort in those locations could be identified through follow-up questionnaires.     

Experimental alveolar echinococcosis in pigs, lesion development and serological follow up

Liver lesions were found in 6/6 pigs 7 months after oral inoculation with 5000 or 35,000 Echinococcus multilocularis eggs. However, lesion morphology differed considerably among the animals. The largest lesions (3–8 mm in diameter) were found in a single pig and smaller lesions (1.5–3 mm) in three pigs. These lesions were clearly circumscribed and had pronounced central necroses and dystrophic calcifications. In contrast, most of the smallest (usually <1.5 mm in diameter) found in two other pigs, had small compact fibrotic areas and blurred borders with obvious fibrous infiltrations into the interlobular tissues. E. multilocularis specific DNA was detected by PCR in all lesion types, but metacestode viability, as assessed by in vivo intraperitoneal inoculations in jirds, could not be demonstrated. Within 1 month post inoculation, all pigs developed specific IgG antibody responses against a battery of different antigens (metacestode, cyst fluid, and protoscoleces-derived native E. multilocularis and E. granulosus antigens, affinity purified Em2G11 antigen, antigen B, recombinant Em II/3–10 antigen). Two different reaction patterns were recorded. In the two pigs with the small lesions, pronounced reactions against all crude antigens with peaks 3–5 months p.i. and clearly elevated levels until the end of the experiment were noted. In all other pigs, antibody reactions remained low in all cases. In conclusion, we demonstrated two types of E. multilocularis metacestode development in pigs with distinct immunological response patterns.