Survey on blood-sucking lice (Phthiraptera: Anoplura) of ruminants and pigs with molecular detection of Anaplasma and Rickettsia spp

Lice may serve as biological or mechanical vectors for various infectious agents. To investigate louse infestation of ruminants and pigs, and pathogens potentially transmitted by them, anopluran lice (n=1182) were collected in Hungary, and evaluated for the presence of anaplasma, rickettsia and haemotropic mycoplasma DNA. On cattle the following species were found: Linognathus vituli (57%), Haematopinus eurysternus (38%) and Solenopotes capillatus (5%). L. vituli had a lower mean individual count/host when compared to H. eurysternus. On calves only L. vituli was observed, with a higher louse burden than on full-grown cattle. H. eurysternus and S. capillatus were more likely to occur simultaneously with another species on the same host, than L. vituli. Goats infested with Linognathus stenopsis had the overall highest prevalence (68%), while pigs harbouring Haematopinus suis showed the lowest (<1%). Anaplasma DNA was detected in 50% of pools analysed. In L. vituli Anaplasma ovis (or a closely related novel Anaplasma marginale genotype) was identified. Anaplasma-positivity of H. suis suggests that pigs may extend the reservoir and/or host spectrum of relevant species. Anaplasma-infected L. stenopsis pools show for the first time that caprine anaplasmosis is endemic in Hungary. Rickettsia spp. were demonstrated from Linognathus spp. and H. eurysternus. No haemotropic mycoplasmas were detected in any samples. In conclusion, this is the first molecularly confirmed report of bovine and ovine Anaplasma spp. in L. vituli, L. stenopsis and H. suis. The present results suggest that phthirapterosis of domestic animals deserves more attention, and lice should be evaluated among the broad range of potential vectors of arthropod-borne pathogens.     

Molecular investigation of transplacental and vector-borne transmission of bovine haemoplasmas

The present study was carried out in a herd with concurrent infections of Mycoplasma wenyonii and 'Candidatus M. haemobos', to investigate if transplacental and/or vector-borne transmission is possible for one or both bovine haemoplasma species. For this purpose blood samples were collected from 38 mother animals and their newborn calves; as well as from 17 uninseminated cows twice three months apart. In addition, 311 mosquitoes and blood-sucking flies (Diptera: Culicidae, Tabanidae, Muscidae) were cought near the animals. DNA was extracted from all samples, followed by real-time PCR analysis. In 10.5% of neonate calves, that were born to cows harbouring both haemoplasmas, M. wenyonii and/or 'Candidatus M. haemobos' positivity was detected. Copy numbers in positive samples from cows and their calves indicated that - in comparison with M. wenyonii - 'Candidatus M. haemobos'-bacteraemia had usually lower levels. In samples of uninseminated cows the rate of infection with the latter species decreased. These findings may explain why M. wenyonii was significantly more frequently detected in blood-sucking flies, than 'Candidatus M. haemobos'. In conclusion, molecular evidence is provided for the first time on the transplacental transmission of bovine haemoplasmas. Regarding their spread by blood-sucking arthropods, new potential vectors were identified, i.e. the horn fly (Haematobia irritans), the stable fly (Stomoxys calcitrans) and two species of horse flies (Tabanus bovinus, T. bromius).     

贵州省山羊流产的病原学鉴定

为了解贵州省山羊流产与山羊痘的相关性,采用琼脂扩散试验和PCR法对本省10个市(县)流产羊群的血清和病料样本进行山羊痘抗原抗体及病原核酸检测,同时血清进行布氏杆菌抗体检测,流产胎儿病料进行羊流产亲衣原体病原核酸检测。结果发现山羊痘羊群流产率达37.1%(4329/11660),山羊痘血清抗体阳性率为38.2%(34/89),抗原阳性率为72.7%(32/44),流产胎儿山羊痘病毒核酸检出率为83.3%(10/12),发病羊群未检出布氏杆菌和羊流产亲衣原体感染。结果表明,山羊流产与山羊痘感染有一定关系,提示在山羊养殖中应加强饲养管理,防止山羊痘感染引起孕羊流产。     

Progressive immunopathological changes during early stages of experimental infection of goats with Mycobacterium avium subspecies paratuberculosis

A dose of 10(10) Mycobacterium avium subspecies paratuberculosis was administered orally on seven occasions to produce experimental paratuberculosis infection in 10 5-8-week-old goat kids. Bacteriological, immunological, and histopathological changes, their relationships, and the efficacy of the commonly used diagnostic methods were studied during the progressive disease up to 270 days postinfection (DPI). Significant lymphocyte proliferative responses in the peripheral blood of five goats were detected as early as 60 DPI. A lymphoproliferative test was also performed on lymphocytes purified from different compartments of the guts of five infected and five control goats. Significant proliferative responses were observed in lymphocytes of jejunal compartments of all five goats, of which four had also significant lymphocyte proliferation in the blood. The ileal lymphocytes from two goats, one each at 120 and 270 DPI, had significant proliferation. The histological lesions were mainly observed in the gut-associated lymphoid tissues of the ileocecal valve, the ileum, and the terminal jejunum. Acid-fast bacilli were demonstrated in the lesions of two goats at 60 and 210 DPI. Bacterial culture showed poor sensitivity, detecting positive results for only one goat in the fecal and tissue samples at 210 DPI, whereas polymerase chain reaction (PCR) detected one goat in fecal sample at 210 DPI and two goats in tissue samples at 60 and 210 DPIs, respectively. Enzyme-linked immunosorbent assay and agar gel immunodiffusion test were found to be 100% sensitive from 180 and 210 DPI onwards, respectively.     

Contents of the glucose transporters GLUT1 and GLUT4 in oxidative and glycolytic muscles of goat kids and adult goats

The objective of this study was to elucidate, whether the impaired insulin sensitivity with regard to glucose utilisation in ruminating goats compared with suckling goat kids may be due to a reduced expression of the glucose transporters GLUT1 and GLUT4 in skeletal muscle. Muscle samples were removed from red, oxidative muscles (M. masseter, diaphragm) and white, glycolytic muscles (M. longissimus dorsi, and M. semitendinosus) of five goat kids fed with milk exchanger and nine adult goats of different stages of life. Samples were analysed for their GLUT1 and GLUT4 contents. The muscles were characterised metabolically by measuring the activities of isocitrate dehydrogenase (ICDH) and of lactate dehydrogenase. In all four analysed muscles the GLUT4 contents of adult goats were significantly (p < 0.05) lower than in goat kids. Significant differences concerning the GLUT4 contents in skeletal muscles were not detected in adult goats of different stages of life. The GLUT1 contents differed to a lower extend between goat kids and adult goats. The results of this investigation indicate that the impaired insulin sensitivity of adult compared with suckling ruminants is accompanied by or leads to a decreased GLUT4 expression.     

河南及重庆、安徽部分地区山羊球虫感染情况与种类调查

采用饱和蔗糖溶液漂浮法对河南省的8个地区、重庆市的云阳县和安徽省的合肥市共1126份山羊粪便样品进行检查,发现1070份球虫阳性样品,总感染率95．03％。对813份阳性样本中的球虫卵囊进行形态学鉴定,共发现12种艾美耳球虫,分别为阿普艾美耳球虫、阿氏艾美耳球虫、艾丽艾美耳球虫、斑点艾美耳球虫、苍白艾美耳球虫、家山羊艾美耳球虫、柯察艾美耳球虫、克氏艾美耳球虫、妮氏艾美耳球虫、山羊艾美耳球虫、羊艾美耳球虫和约奇艾美耳球虫。山羊最多可同时感染9种球虫,多数为2-4种,混合感染率78．7％;6月龄以下、6-12月龄和1岁以上山羊球虫感染率分别为93．3％、97．1％和96-3％,平均OPG值分别为5282．77、3550．71和1507．88;除奶山羊外,不同品种山羊球虫感染无明显差异;舍饲和放牧山羊球虫感染率分别为95．7％和92．8％,平均OPC值分别为3744．35和1028．62。跟踪调查显示,舍饲山羊球虫感染无明显季节性,但夏秋季节球虫感染强度高于冬春季节。     

In vivo and in vitro propagation and transmission of Toggenburg orbivirus

The Toggenburg orbivirus (TOV), a recently discovered virus related to bluetongue virus (BTV), has been identified in goats in Switzerland, Italy and Germany. Isolation of TOV in vitro has not yet been achieved and the transmission mechanisms are still unknown. In the experimental infection of pregnant goats described here, TOV could not be detected in secretion/excretion samples or fetal blood. Material from the goat experiment was used as inoculum for propagating the virus in vitro. To enhance the infectivity of TOV several modified protocols, e.g. pretreatment of the virus with trypsin, polyethylene glycol-mediated infection and lipofection were applied. Isolation of TOV, attempts to infect Culicoides nubeculosus by feeding TOV-positive blood and intracerebral inoculation of newborn mice were unsuccessful. The results of these studies suggest that TOV requires specific but different factors than other BTVs for infection and replication outside of its natural caprine host.     

Molecular investigation,isolation and phylogenetic analsysis of Coxiella burnetii from aborted fetus and ticks

Q fever is a zoonotic infection threatening human health, causing abortions in cattle, sheep and goats. Coxiella burnetii (C. burnetii) also causes serious problems such as low birth weight, infertility. This study is the first exemplary for analysis of Q fever around Black Sea region in Turkey. In the study, a total of 270 aborted fetuses (171 cattle, 79 sheep, 20 goats) and 1069 tick samples were aimed to be searched by PCR method. C. burnetii DNA was detected in 8 (2.96 %) of 270 sheep specimens while it could not be found in cattle and goat specimens. 406 sample pools were created from 1069 tick samples (490 male, 579 female) collected from 254 farm animals (187 cattle, 54 sheep, 13 goats) and 11 of these were stated positive. Tick species determined as C. burnetii positive were Hyalomma marginatum, Hyalomma anoliticum excavatum, Hyalomma detritum and Boophilus annulatus. Agent isolation was carried out within embryonated eggs. Agents were stained with Giemsa and was showed. Sequence analysis was performed for TUR/SAM/coxiella_1 (MN917207) isolate and phylogenetic tree was created. This tree, created in compliance with IS1111 transposon gene, did not form different branches in regard to host affiliation (goat, sheep, tick, human) and geographical distribution. As a result, an important zoonotic agent, C. burnetii was diagnosed in sheep aborted fetuses and the infection was proved to have spread among sheep herds in Black Sea region. Besides, 4 separate tick species found in our region hosted the agent and were found important for infection.     

IS900 restriction fragment length polymorphism (RFLP) analysis of Mycobacterium avium subsp. paratuberculosis isolates from goats and cattle in Norway

In Norway, paratuberculosis has been frequently diagnosed in goats, while cattle have been almost free of the infection. This difference in prevalence between goats and cattle has led to speculations about the existence of a Mycobacterium avium subsp. paratuberculosis (M. a. paratuberculosis) isolate that is non-pathogenic for cattle. There is little information available on genotypic variation of M. a. paratuberculosis isolated from animals in Norway. In the present study, genotypic information on 51 isolates from goats and four isolates from cattle in Norway was obtained by use of IS900 restriction fragment length polymorphism (RFLP) analysis. All isolates from cattle and 84% of the isolates from goats had the same RFLP pattern (B-C1). Five RFLP patterns not previously detected were found. No genotypic variation that could explain a difference in host origin was found between the isolates from cattle and the majority of the Norwegian goat isolates. This lack of difference indicates that the most common M. a. paratuberculosis isolates in Norway may infect both cattle and goats.     

Pathogenicity of different species of staphylococci in caprine udder

Aseptic foremilk samples were collected from Finnish landrace goats. Ten different species of staphylococci, causing subclinical infections were detected. Twelve goats with persistent subclinical staphylococcal infection were followed on a monthly basis and compared with foremilk samples of nine goats suffering from clinical mastitis. Parameters of inflammation based on the activity of the California Mastitis Test (CMT). N-acetyl-beta-D-glucosaminidase (NAGase) and antitrypsin were determined from the milk. Staphylococci were further classified using the API STAPH system. On the basis of elevations of activities of CMT, NAGase and antitrypsin, Staphylococcus aureus was the most pathogenic in clinical and subclinical mastitis, S. hyicus showed only marginal pathogenicity. Subclinical infections were persistent and the infective organism was not always detected from milk by culture. The biochemical reactions of subclinical staphylococci seemed to vary within the same gland by time. Antitrypsin was most effective in differentiating between subclinical and clinical infection. A teat cistern puncture technique was found to be suitable for the goat.     

Use of scanning electron microscopy to confirm the identity of lice infesting communally grazed goat herds

Lice have been described on goats in commercial farming systems in South Africa but not from flocks on communal grazing. During a longitudinal survey on the causes of goat kid mortality, conducted in Jericho district, North West Province, lice were collected from communally grazed indigenous goats. These lice were prepared for and viewed by scanning electron microscopy, and micro-morphological taxonomic details are described. Three species of lice were found in the study area and identified as Bovicola caprae, Bovicola limbatus and Linognathus africanus. Sucking and biting lice were found in ten of the 12 herds of goats examined. Lice were found on both mature goats and kids. Bovicola caprae and L. africanus were the most common biting and sucking lice respectively in all herds examined. Scanning electron microscopy revealed additional features which aided in the identification of the louse species. Photomicrographs were more accurate aids to identification than the line drawings in the literature and facilitated identification using dissecting microscope.     

基于SNP芯片的云上黑山羊遗传结构分析

为从分子水平对云上黑山羊遗传结构做出评价,本研究以云上黑山羊核心群母羊108只(来自7个育种核心场、10个群体)、育种素材云岭黑山羊和努比山羊各11和14只、对照组波尔山羊10只为试验动物,用Illumina公司Iselect Goat60k芯片技术在全基因组范围内进行单核苷酸多态性(SNP)分析,并运用GenAlEx 6.5、Cervus 3.0、SNPRelate、Plink 1.07、Mega 4.0进行遗传多样性参数统计计算、主成分分析和系统进化树构建。试验获得143个样本在48 116个SNPs位点上的分型结果,其中波尔山羊10个样本单独聚集,与云上黑山羊、努比山羊、云岭黑山羊存在较大的遗传距离,较小的遗传一致度和较小的基因流,在聚类进化树中显示为一个独立分支,这些结果与波尔山羊实际遗传背景完全一致,显示出它在本研究中的对照组作用,同时证明本研究方法可行、结果可靠;云上黑山羊108个样本具有高度一致性,在聚类系统树中这些样本聚为一个大簇,而努比山羊(14个)和云岭黑山羊(11个)的样本各自聚为一簇;UPGMA进化树显示,云上黑山羊与努比山羊关系较近,与云岭黑山羊关系稍远,这与云上黑山羊育种导血方案结果相一致。云上黑山羊新品种在基因组水平上可与其育种素材明显区分,在基因组层面已具备独立品种特点。     

In vivo and in vitro propagation and transmission of Toggenburg orbivirus

The Toggenburg orbivirus (TOV), a recently discovered virus related to bluetongue virus (BTV), has been identified in goats in Switzerland, Italy and Germany. Isolation of TOV in vitro has not yet been achieved and the transmission mechanisms are still unknown. In the experimental infection of pregnant goats described here, TOV could not be detected in secretion/excretion samples or fetal blood. Material from the goat experiment was used as inoculum for propagating the virus in vitro. To enhance the infectivity of TOV several modified protocols, e.g. pretreatment of the virus with trypsin, polyethylene glycol-mediated infection and lipofection were applied. Isolation of TOV, attempts to infect Culicoides nubeculosus by feeding TOV-positive blood and intracerebral inoculation of newborn mice were unsuccessful. The results of these studies suggest that TOV requires specific but different factors than other BTVs for infection and replication outside of its natural caprine host.     

Toggenburg Orbivirus, a new bluetongue virus: Initial detection, first observations in field and experimental infection of goats and sheep

A novel bluetongue virus termed “Toggenburg Orbivirus” (TOV) was detected in two Swiss goat flocks. This orbivirus was characterized by sequencing of 7 of its 10 viral genome segments. The sequencing data revealed that this virus is likely to represent a new serotype of bluetongue virus [Hofmann, M.A., Renzullo, S., Mader, M., Chaignat, V., Worwa, G., Thuer, B., 2008b. Genetic characterization of Toggenburg Orbivirus (TOV) as a tentative 25th serotype of bluetongue virus, detected in goats from Switzerland. Emerg. Infect. Dis. 14, 1855–1861].In the field, no clinical signs were observed in TOV-infected adult goats; however, several stillborn and weak born kids were reported. Although born during a period of extremely low vector activity, one of these kids was found to be antibody and viral genome positive and died 3.5 weeks postpartum.Experimental infection of goats and sheep, using TOV-positive field blood samples, was performed to assess the pathogenicity of this virus.Goats did not show any clinical or pathological signs, whereas in sheep mild bluetongue-like clinical signs were observed. Necropsy of sheep demonstrated bluetongue-typical hemorrhages in the wall of the pulmonary artery. Viral RNA was detected in organs, e.g. spleen, palatine tonsils, lung and several lymph nodes of three experimentally infected animals.Unlike other bluetongue virus serotypes, it was not possible to propagate the virus, either from naturally or experimentally infected animals in any of the tested mammalian or insect cell lines or in embryonated chicken eggs.In small ruminants, TOV leads to mild bluetongue-like symptoms. Further investigations about prevalence of this virus are needed to increase the knowledge on its epidemiology.     

Experimental studies on the pathogenicity of Mycoplasma ovipneumoniae and Mycoplasma arginini for the respiratory tract of goats.

Mycoplasma ovipneumoniae and Mycoplasma arginini were the species of Mollicutes most commonly isolated from 175 goats with respiratory disease in Ontario. The pathogenicity of M. ovipneumoniae, strain B321B and M. arginini, strain D53e, was assessed in goats following endobronchial inoculation. One out of three two year old goats developed fever after inoculation with a pure culture of strain B321B, and it had extensive subacute fibrinous pleuritis when necropsied three weeks later. Neither of the remaining goats had lesions in the respiratory tract. Mycoplasma ovipneumoniae was recovered from one of the animals four days after inoculation, but not at necropsy from any of the goats, at which time a marked humoral immune response with growth inhibiting antibodies was detected. In a second experiment three four to five week old goats were inoculated with the same strain and three other goats were given placebo treatment. One experimental goat developed fever and coughing, and it had extensive subacute fibrinous pleuritis in the right side and pneumonia. Another goat had focal pneumonia in the left diaphragmatic lobe. Microscopically there was subacute hyperplastic suppurative bronchiolitis, atelectasis and nonsuppurative alveolitis. The infected animals did not clear the mycoplasma and not all of them produced antibodies. Mycoplasma arginini, strain D53e, did not induce lesions in any of four goat kids within 14 days after inoculation but did cause transient elevations in rectal temperature, circulating monocytes, circulating neutrophils and blood fibrinogen. Mycoplasma arginini was infective and immunogenic for all inoculated animals and showed a particular affinity for the tonsil. Thus, this study provides the first evidence that M. ovipneumoniae is pathogenic for goats causing pneumonia and pleuritis.(ABSTRACT TRUNCATED AT 250 WORDS)     

Experimental contagious caprine pleuropneumonia: a long term study on the course of infection and pathology in a flock of goats infected with Mycoplasma capricolum subsp. capripneumoniae

Contagious caprine pleuropneumonia (CCPP) is a major threat to goat farming in parts of Africa and Asia. It classically causes acute high morbidity and mortality early in infection, but little is known of its long term epizootiology and course. In this study, 10 goats were inoculated with Mycoplasma capricolum subsp. capripneumoniae (M. capripneumoniae) and then mixed with 15 goats for contact transmission. The disease course was monitored in each goat for 56-105 days, whereafter the goats were killed and necropsied. Varying features signifying infection occurred in altogether 17 goats (7 inoculated, 10 in-contact). Clinical signs were severe in 8 goats but no fatalities occurred. Only 6 goats had serum antibody titres against M. capripneumoniae in ELISA. Fourteen goats (5 inoculated, 9 in-contact) had chronic pleuropulmonary lesions compatible with CCPP at necropsy and 7 of those showed M. capripneumoniae antigen in the lung by immunohistochemistry. Neither cultivation nor PCR tests were positive for the agent in any goat. The results indicate that the clinical course of CCPP in a flock may be comparatively mild, M. capripneumoniae-associated lung lesions may be present at a late stage of infection, and chronic infection may occur without a significant serological response.     

The seroprevalence of Japanese encephalitis virus in goats raised in Korea

Japanese encephalitis virus (JEV) causes a mosquito-borne viral zoonosis that is becoming increasingly important to public health in east and south Asia. Although JEV is primarily associated with reproductive failure in swine, JEV infection can cause fever and headache in humans and is associated with aseptic meningitis and encephalitis. The exact mode of transmission, including host range and possible source of viral amplification within livestock, is still not completely clear. This study consisted of a serological survey of JEV infection in goats. A total of 804 goat serum samples were collected from 144 farms in Korea between May 2005 and May 2006. The incidence of positive cases was 12.1% (97 out of 804 goats). The seroprevalence of JEV infection in the 144 farms screened was 31.3% (45/144), indicating that JEV infection is frequent in goat farms in Korea. In addition, three districts of Korea (mainly in the southern region) had a higher seroprevalence of JEV compared to other areas. The results suggest that goats could be monitored epidemiologically as a sentinel animal for JEV transmission in Korea.     

Diagnosis of paratuberculosis by fecal culture and ELISA on milk and serum samples in two types of Chilean dairy goat herds.

Fecal culture has been the primary method used to diagnose paratuberculosis in goats. It is laborious, slow, and expensive. Validation of enzyme-linked immunosorbent assays (ELISAs) on milk samples could make paratuberculosis testing more widely available for goat farmers. The aim of this study was to determine the accuracy of serum and milk ELISAs for paratuberculosis, relative to fecal culture, in Chilean dairy goats. Eight dairy goat herds were selected. Feces, blood, and milk samples were collected from all female goats >2 years old. Fecal samples were cultured using Herrold egg yolk medium with mycobactin J and antibiotics. Serum and milk samples were tested using a commercial ELISA kit for Mycobacterium avium subsp. paratuberculosis antibody detection. A total of 383 goats were tested by ELISA and fecal culture. The sensitivity of ELISA on serum and milk relative to fecal culture was 74.3% (95% CI: 59.8-88.8) and 60% (95% CI: 43.8-76.2), respectively. The corresponding values for ELISA specificity based on the percentage of non- M. avium subsp. paratuberculosis-infected goats testing ELISA-negative were 98.6% (95% CI: 96.6-100) and 99.3% (95% CI: 97.9-100) on serum and milk, respectively. Proportions of positive results for serum and fecal samples were significantly different, whereas the proportions of positive results for milk and fecal samples were not significantly different. The milk ELISA had a moderate level of agreement with fecal culture results (Kappa = 0.57). The paratuberculosis ELISA on goat milk samples may be a cost-effective, accurate alternative to fecal culture.     

陕西省某羊场简阳大耳山羊球虫感染情况调查及种类鉴定

为了解陕西省某简阳大耳山羊场球虫种类和感染情况,采集不同年龄段简阳大耳山羊新鲜粪便55份,用形态学方法并结合麦克马斯特虫卵计数法(McMaster's method)对该场简阳大耳山羊球虫进行种类鉴定和感染情况调查。结果表明,大耳山羊球虫总感染率为92.7%,其中3月龄~7月龄、10月龄~12月龄、3岁山羊球虫检出率分别为100%、90%和86.7%,各年龄段平均OPG值为3 300、1 050、854。共鉴定出8种山羊球虫,其中阿氏艾美耳球虫(Eimeria arloingi)、雅氏艾美耳球虫(E.ninakohlyakimovae)和艾氏艾美耳球虫(E.alijevi)为该羊场优势虫种。