Morphological survey of bovine Setaria in the abdominal cavities of cattle in Aomori and Kumamoto Prefectures, Japan

An epidemiological survey of bovine Setaria collected from the abdominal cavities was performed morphologically on the cattle in Aomori and Kumamoto Prefectures, Japan, between August 2005 and July 2006. Fifty Setaria worms were collected from the cattle in Aomori Prefecture and 847 from those in Kumamoto Prefecture. Of these worms, 35 were identified as Setaria digitata, 14 as S. marshalli, and one as S. labiatopapillosa in Aomori Prefecture, while 816 were identified as S. digitata and 31 as S. marshalli in Kumamoto Prefecture.     

Demonstration of vector competence of Culex quinquefasciatus (Diptera: Culicidae) for Setaria digitata

In Taiwan, Setaria digitata infection causes a lumber paralysis in increasing number of cattle. Culex quinquefasciatus is one of the predominant mosquitoes, and it has been suspected that C. quinquefasciatus acts as a vector to Setaria nematodes prevalence but this was not confirmed. C. quinquefasciatus, Aedes albopictus and A. aegypti of various strains were investigated using an artificial infection system to evaluate their vector competence. After blood feeding at day 14, the number of larvae (stage III) per infected mosquito in A. aegypti (Liverpool strain), A. aegypti (Kaohsiung strain), A. aegypti (Tungan strain), C. quinquefasciatus (Taichung strain) and A. albopictus (Taichung strain) was 1.3 +/- 0.1, 1.3 +/- 0.1, 1.4 +/- 0.1, 1.0 +/- 0.0 and 0 +/- 0.0 (mean +/- S.E.M), respectively. The vector efficiency index of A. aegypti (Liverpool) was the highest among mosquitoes whereas A. albopictus showed a complete refractoriness to the infection. In conclusion, C. quinquefasciatus demonstrates its potential competence for serving as a transmission vector of S. digitata. This mosquito might therefore be responsible, at least in part, for the prevalence of cattle lumbar paralysis in Taiwan. This is the first report of C. quinquefasciatu demonstrating its vector competence for S. digitata.     

Setaria digitata in cattle of Thailand identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis.

Adult Thai Setaria worms collected from cattle which were bred, housed and slaughtered in Thailand were morphologically identified as Setaria digitata. Furthermore, in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) adult Thai S. digitata had the same protein profiles as adult Japanese S. digitata, but did not possess the protein with a molecular size of 69 kDa which was confirmed in adult S. marshalli. In addition, there were no differences in the protein profiles between male and female S. digitata. In point of the distribution pattern of the proteins ranging from 73 to 64 kDa revealed by 2D-PAGE, there were no differences between Thai and Japanese S. digitata, and between male and female worms of the species.     

辽宁庄河奶山羊“腰麻痹病”病原学及病理形态学研究

5例自然感染“腰麻痹病”奶山羊的病灶均局限在脑脊髓系统,其中1例脊髓腔中发现1条活泼的指状腹腔丝虫幼虫,在另一例脊髓实质病变部发现了与指状腹腔丝虫断面相似的虫体断面.各例病变性质均属虫伤性液化坏死性非化脓性脑脊髓炎变化.由此认为,指状腹腔丝虫幼虫是本病的病原.病灶的超微结构观察表明,病变无特异性,主要呈退行性病理变化:神经元、神经胶质细胞及细胞器呈严重的细胞病变,有髓神经纤维脱髓鞘,无髓神经纤维变性溶解,神经毡形成坏死性空洞及出血等变化.     

Ocular infection of cattle with Setaria digitata

One 5-month-old female native Korean calf and a 2-year-old female Holstein cow raised in two farms about 4 km apart from each other in Korea, were found to have the left eye opaque, which included motile white worms in the aqueous humor. The parasite removed from the left eye of the calf was identified as Setaria digitata based on both light and electron microscopic features. The ocular infection with S. digitata reported herein may document the first aberrant case in Korean cattle.     

牛腹腔丝虫对3种伊蚊的感染试验

用牛腹腔指状丝虫和唇乳突丝虫的微丝蚴感染埃及伊蚊、白纹伊蚊和东乡伊蚊,观察了蚊虫的感染情况及牛腹腔丝虫Ⅲ期蚴的形态。结果,这两种丝虫的微丝蚴在3种伊蚊体内都能发育成熟,各蚊种的感染率不同,埃及伊蚊和白纹伊蚊分别为40%和30%以上,东乡伊蚊近70%;Ⅲ期蚴的表面结构不因蚊种而异。以不同蚊体内发育而成的Ⅲ期蚴感染小鼠,均可致小鼠出现与马、羊脑脊髓丝虫病相似的瘫痪或转圈运动等神经症状。由此认为,埃及伊蚊和白纹伊蚊也可能为脑脊髓丝虫病的传播媒介。     

Comparison of microfilaria concentration method for Setaria digitata infection in cattle and for Dirofilaria immitis infection in dogs

Several peripheral blood microfilaria concentration methods that use Acetone (Acetone test), 2% formalin (modified Knott method), 5% Tween 20 solution, distilled water, 1% or 0.1% SDS were compared for their efficacy in detecting Setaria digitata microfilaria in cattle. The Acetone test was found to be more efficacious than the modified Knott method or the 5% Tween 20 solution test for detecting the S. digitata microfilaria in bovine blood. However, besides the Acetone test, the modified Knott method was also found to be suitable for Dirofilaria immitis microfilaria detection in dogs. SDS and distilled water were found not to be effective as hemolytic agent for the disruption of the red blood cell of both the cattle and dogs. Thus, the Acetone test is recommended for the primary screening of microfilaremia of S. digitata in cattle.     

Patent infection with Setaria digitata in goats in Saudi Arabia.

A survey was carried out on Saudi sheep and goats during 1995, at the Veterinary Diagnostic laboratory, Jeddah, to study Setaria infection in small ruminants in Saudi Arabia. Setaria digitata worms were detected in the abdominal cavity of 5 out of 48 goats (10.4%). Sheep were not found to be infected. All infected goats were adults of 2-4 years of age and originating from the Jeddah, Mecca and Al-Leith areas. Four cases were recorded during January-March and one during August. No lesions due to S. digitata were found in the visceral organs and brains of the infected goats. This is the first record of patent infection with S. digitata in goats.     

A sensitive polymerase chain reaction based assay for the detection of Setaria digitata: the causative organism of cerebrospinal nematodiasis in goats, sheep and horses

A sensitive PCR assay for the detection of Setaria digitata has been developed. Two oligonucleotide primers (17 nt) were designed from a previously cloned and characterized tandemly arranged repetitive sequence of Setaria digitata. Using these primers, it was possible to amplify small quantities (100 fg) of S. digitata genomic DNA. A simple procedure, using proteinase K and non-ionic detergent NP 40, was followed to process the host blood samples and mosquitoes harbouring L3 larvae. The sensitivity of the polymerase chain reaction based assay surpasses the microscopic detection and the previously reported oligonucleotide based chemiluminescent detection of microfilariae in infected host blood samples and L3 larvae in mosquitoes.     

Differentiation of Setaria digitata and Setaria labiatopapillosa using molecular markers

5S rRNA intergenic regions of Setaria digitata and Setaria labiatopapillosa were PCR amplified with primers designed from the 5S rRNA gene of Brugia malayi. The ladder-like banding patterns obtained for the amplifications were distinctly different for the two species. Four amplified products were cloned into the pBS vector and completely sequenced. DNA clones from two individual samples of S. digitata, Sd4 and Sd6, showed 97% sequence homology to each other. All sequenced clones showed the presence of the spliced leader (SL) RNA gene with a 22 nucleotide spliced leader sequence. The phylogenetic tree constructed using these data and the 5S rRNA intergenic regions of several other filarial nematodes showed the Setaria species sharing a branch with Dirofilaria. RAPD-PCR analyses identified 107 bands of which 86 were polymorphic (80%). A dendrogram constructed for S. digitata and S. labiatopapillosa separated the two species into two distinct clusters. The polymorphic loci identified by the RAPD-PCR analyses can be studied further to develop species-specific probes/PCR primers for the identification of each species.     

唇乳突丝虫感染蚴接种非固有宿主动物致发脑脊髓丝虫病研究

用东乡伊蚊叮吮含有唇乳突丝虫微丝蚴的模型动物——小鼠血液,待微丝蚴在蚊体内发育至感染期时,分离出感染坳,皮下多点接种43只昆明小鼠(每只15～200条)、1只山羊及2只绵羊(每只200～300条)和4匹驹(每匹750～1 250条);将指状丝虫接种到20只昆明鼠体内,比较两种虫体的致病性。结果:唇乳突丝虫感染鼠有16只经1～8 d潜伏期发病,呈现瘫痪、昏迷症状后死亡.实验羊和驹经5～21d潜伏期后均呈不同程度的运动和神经症状,于接种虫体后40～140 d扑杀.经病理学观察,3种动物脑脊髓均呈现虫伤性液化坏死灶及非化脓性脑脊髓炎变化,在其中枢神经系统组织切片中发现丝虫虫体断面或钙化碎片,从而证明唇乳突丝虫可人工感染昆明小鼠、羊、驹发生脑脊髓丝虫病,进而提出该虫可以成为马、羊脑脊髓丝虫病的病原之一。指状丝虫感染鼠死亡率高于唇乳突丝虫感染鼠(P<0.01),初步认为唇乳突丝虫致病性较指状丝虫弱。     

Absence of Wolbachia endobacteria in Sri Lankan isolates of the nematode parasite of animals Setaria digitata

Setaria digitata is an animal filarial parasite with natural hosts of cattle and buffaloes that causes mild disease conditions. Infection of non-permissive hosts such as goats, sheep and horses, by this nematode can cause cerebrospinal nematodiasis that leads to lumbar paralysis and the eventual death of the animals and inflicts considerable economic losses on livestock farmers. Wolbachia are obligate mutualistic endosymbionts for some filarial nematodes and are currently being targeted for the control of diseases caused by these parasites. However, little is known about the occurrence of this endosymbiont in the Setariidae family. In this work, worms collected from infected cattle in Sri Lanka were morphologically identified as S. digitata and tested for the presence of Wolbachia by PCR screening using the WSP- and Wolbachia-specific 16S rRNA and multilocus sequence typing primers that were designed to amplify the gatB, coxA, hcpA, ftsZ and fbpA sequences of Wolbachia. The presence of endobacteria in S. digitata was also examined by whole-mount immunofluorescence staining of the parasites and transmission electron microscopic studies. These analyses did not produce evidence of presence of Wolbachia or any other endosymbiotic bacteria in S. digitata, whereas such evidence was found in Brugia malayi, which was used as a positive control in this study.     

Inflammatory cytokines and antigen-responsive mononuclear cells in peripheral blood of cattle infected with Salmonella takoradi

To determine the immunological response in lactating dairy cows infected with Salmonella (S.) Takoradi, the relationships among distributions of peripheral blood mononuclear cell (PBMC) subpopulations, endotoxin concentrations and dynamics of inflammatory cytokines in blood were investigated. The ratio of CD4+ T cells to CD8+ T cells was significantly lower in the affected cattle than in the control cattle (p<0.05) to decrease in the number of CD4+ T cells in the infected cattle. In contrast, the numbers of gammadeltaT cells, MHC class II-positive cells were significantly higher in the affected cattle than in the control cattle (p<0.01 respectively). Endotoxemia was found in all but one of the affected cattle. Serum IL-1 and IL-6 bioactivities were significantly higher in the affected cattle than in the control cattle (IL-1, p<0.05; IL-6. p<0.01). Serum TNF-alpha activities and levels were not detected in the control and affected cattle. The activities of proinflammatory cytokines determined by the bioassay are important to the relationships between concentration of endotoxin, cytokines and clinical signs. such as leukocytosis, leukopenia, fever or bacterial shedding. Serum IL-2 levels were lower in the affected cattle than in the control cattle. Serum IFN-y was not detected in the affected cattle except one. These results by the ELISA seemed to reflect the condition of subpopulation in the PBMCs from the shedding cattle. The present results suggest that cellular immunity is suppressed while the humoral immunity is activated in acute bovine salmonellosis.     

Anaplastic astrocytoma in the spinal cord of an African pygmy hedgehog (Atelerix albiventris)

A 2-year-old, female hedgehog presented with an 8-month history of progressive, ascending paresis/paralysis and was tentatively diagnosed with wobbly hedgehog syndrome. She died awaiting further diagnostic tests, and the owners consented to postmortem examination. Grossly, the bladder was large and flaccid and the cervical and lumbar spinal cord were regionally enlarged, light grey, and friable with multifocal hemorrhages. The thoracic spinal cord was grossly normal. Microscopically all regions of the spinal cord had similar changes, although the cervical and lumbar sections were most severely affected. These regions were completely effaced by a moderately cellular infiltration of highly pleomorphic polygonal to spindle shaped cells, mineralization, and necrosis, which were most consistent with anaplastic astrocytoma. The thoracic spinal cord white matter was similarly infiltrated by the neoplastic cells, with perivascular extension into the otherwise normal grey matter. A diagnosis of anaplastic astrocytoma was confirmed using immunohistochemical stains that were positive for glial fibrillary acidic protein and S100.     

Cryopreservation of some common nematodes of ruminants for up to 11,3 years

Infective larvae of selected batched of the following nematode species from sheep and cattle were examined for survival and infectivity (by injection into either the abomasum, the duodenum, or the jugular vein) after having been stored in liquid nitrogen for 103-136 months: Haemonchus contortus, Haemonchus placei, Ostertagia circumcincta, Ostertagia ostertagi, Marshallagia marshalli, Cooperia spp., Trichostrongylus axei, Trichostrongylus colubriformis, Trichostrongylus falculatus, Nematodirus spathiger, Nematodirus helvetianus, Oesophagostomum columbianum, Oesophagostomum radiatum, Chabertia ovina and Dictyocaulus filaria. Excluding D. filaria, a mean of 97.7% of the ovine and 96.0% of the bovine nematode larvae were alive when thawed. Compared with previous investigations in this series, little or no reduction occurred with time in either the survival or the viability of the nematodes from cattle, or in the survival of those from sheep. In contrast, the larvae developed poorly in sheep, possibly owing to parenteral treatment of the animals with ivermectin at a dosage of 0.4 mg kg-1, either 6 or 8 days before they were infected.     

Postural and neurological deficits in broiler chicks after cervical vaccination with live vaccine.

A disease characterized by paresis and paralysis was seen in 7-9-day-old broiler chicks after vaccination in the neck area at day-of-age with a live virus vaccine containing viruses of Marek's disease, fowl pox, and infectious bursal disease. Affected birds presented with variable signs of ataxia, lateral recumbency, leg paralysis, and twisting or S-shaped flexure of the neck. Gross lesions noted at necropsy included swelling and edema of the subcutaneous tissues and muscles of the neck at the injection site area. A heavy mononuclear inflammatory cell infiltration was seen in the subcutaneous tissues, connective tissues, and muscles of the neck at the injection site. In some cases, the inflammatory process extended along fascial planes to involve the epidural spaces surrounding the spinal cord. Fatty changes with possible demyelination of nerve fibers were noted in some sections of the spinal cord adjacent to the inflammatory lesions. Clusters of poxviruses were found within some inflammatory lesions on transmission electron photomicrographs.     

Type C botulism in young dogs

A diffuse lower motor neurone paralysis developed in a 6-month-old male Australian cattle dog pup 4 days after it had eaten the carcase of a rotting duck in Centennial Park, Sydney. Two other dogs which ate smaller portions of the same carcase were less severely affected. Clostridium botulinum type C was isolated from and C. botulinum type C toxin was detected in faeces from the severely affected dog. The serum contained 25 LD50 of toxin/ml. The high C. botulinum count and toxin level in the faeces declined progressively during the ensuing weeks, but 114 days after ingesting the carcase C. botulinum type C was still present in faeces and a low toxin titre persisted. Soil, mud and water samples in the area of the duck ponds in the park contained C. botulinum type C spores. Spores and high toxin titres were also found in the intestine of the carcases of 2 birds in the area.     

Proliferative potential of a spinal nephroblastoma in a young dog

The proliferative potential of a spinal nephroblastoma was studied in a young dog. A 4-month-old, female golden retriever showed developing deterioration in her gait and subsequent paralysis of her hind legs. At necropsy, a well-demarcated grayish brown tumor mass was found in the lumbar spinal cord segments between L2 and L3. Histologically, a blastemal cell tumor with a tubule- or glomeruli-like structure was found to be infiltrating intradurally. Proliferating cells at the S-phase, assessed using the bromodeoxyuridine (BrdU) labeling method, were seen occasionally in the tubular cells and glomeruli-like structures and were frequently seen in the blastemal cells. Immunohistochemically, the tubular epithelial cells were positive for cytokeratin, and the blastemal cells were positive for vimentin. The present tumor showed a high potential for growth and invasion, which suggests that it the potential to expand into the adjacent spinal cord.     

Cryopreservation of the infective larvae of the common nematodes of ruminants

Exsheathed infective larvae (L 3) of 19 species of nematodes were tested for infectivity in either sheep or cattle after they had been frozen in 0,9% NaCl solution, stored for a relatively short time in the gas phase of liquid nitrogen and subsequently thawed. In addition, 13 of these species were tested after similar storage for up to 18 months. In sheep, Haemonchus contortus, Ostertagia circumcincta, Trichostrongylus axei, Trichostrongylus colubriformis, Nematodirus spathiger and Oesophagostomum columbianum were viable after 2 years of cryopreservation, a mean of greater than 90% of the L 3 being alive when thawed after this period. Similar results were obtained with Chabertia ovina L 3 after 18 months and with Marshallagia marshalli, Trichostrongylus falculatus and Dictyocaulus filaria, after a short period of freezing. On the other hand, Gaigeria pachyscelis and Strongyloides papillosus survived freezing for up to 7 months but neither was viable at the end of this period, nor was exsheathed G. pachyscelis viable without freezing. Most of these infestations were established by inoculating the infective larvae into the abomasum and/or duodenum. M. marshalli, T. falculatus and C. ovina also proved infective after oral dosing. D. filaria, the only other species tested by this route, was not infective when dosed per os after thawing. The infective larvae of the bovine nematodes, Haemonchus placei, Ostertagia ostertagi, Nematodirus helvetianus, Oesophagostomum radiatum, Cooperia pectinata and Cooperia punctata survived freezing for a mean of 26 months, greater than 90% being alive on thawing, but infectivity was generally lower than with the same genera in sheep. Even when not frozen, exsheathed Bunostomum phlebotomum was non-infective. When Cooperia spp. after thawing were tested for infectivity by the oral route, more worms developed in one calf infested orally than in another infested by inoculation into the duodenum. Ova of H. contortus, M. marshalli, O. circumcincta, T. colubriformis, T. falculatus, N. spathiger, C. ovina, H. placei, O. ostertagi, Cooperia spp. and N. helvetianus were recovered from the faeces of animals infested with cryopreserved L 3. No ova of O. columbianum or O. radiatum were recovered from faeces, because differential larval counts were performed before they were patent. Nevertheless, gravid females were obtained post-mortem. Frozen L 3 of N. helvetianus were used to re-establish a pure strain in calves, 2,3 million ova being recovered from infestations with 10 670 L 3 frozen for 26 months. The infectivity of the progeny of frozen L 3 was tested with M. marshalli and C. ovina. In both instances infectivity was high and the worms which developed also produced ova, thus completing the cycle. This appears to be the first report of infective larvae of parasitic nematodes retaining their infectivity after being frozen in liquid nitrogen (gas phase) for longer than 2 years. This is also apparently the first time that M. marshalli T. colubriformis, T. falculatus, T. axei, N. spathiger, C...