Early and Pre-Clinical Detection of Prion Seeding Activity in Cerebrospinal Fluid of Goats using Real-Time Quaking-Induced Conversion Assay.

Since 2005, two cases of natural bovine spongiform encephalopathies (BSE) have been reported in goats. Furthermore, experimental transmissions of classical (C-BSE) and atypical (L-BSE) forms of BSE in goats were also reported. To minimize further spreading of prion diseases in small ruminants the development of a highly sensitive and specific test for ante-mortem detection of infected animals would be of great value. Recent studies reported high diagnostic value of a second generation of cerebrospinal fluid (CSF) Real-Time Quaking-Induced Conversion (RT-QuIC) assay across a wide spectrum of human prions. Here, we applied this improved QuIC (IQ-CSF) for highly efficient detection of TSEs prion protein in goat cerebrospinal fluid. IQ-CSF sensitivity and specificity were evaluated on CSF samples collected at disease endpoint from goats naturally and experimentally infected with scrapie or bovine isolates of C-BSE and L-BSE, respectively. Next, CSF samples collected from L-BSE infected goats during pre-symptomatic stage were also analysed. PrPL-BSE associated seeding activity was detected at early time points after experimental inoculation, with an average time of 439 days before clinical symptoms appeared. Taken together these data are indicative of the great potential of this in vitro prion amplification assay as ante-mortem TSE test for live and asymptomatic small ruminants.

Diagnostics and Ancillary Tests of Neurologic Dysfunction in the Ruminant.

A variety of diagnostic tests can be used to help further characterize and diagnose neurologic disease in ruminant species. Cerebrospinal fluid is easily collected, and analysis can help in defining the broad category of disease. Diagnostic imaging, including radiography, myelography, ultrasonography, computed tomography, and MRI, have all been used to varying degrees in ruminants. Advanced cross-sectional imaging techniques have the capacity to aid greatly in diagnosis, but their cost can often be prohibitive. Currently, electrodiagnostic tests are not well evaluated or used in the diagnosis of neurologic disease in ruminants.

Application of polymerase chain reaction on cerebrospinal fluid for diagnosis of cerebral coenurosis in small ruminants.

Sheep and goats serve as intermediate hosts for the canine tapeworm Taenia multiceps. The cysts produced by the intermediate stage of parasite are usually found in the cerebral hemispheres of small ruminants, and the resulting disease is commonly known as coenurosis. Coenurosis is clinically manifested in the form of various nervous symptoms, depending on the exact location of the cyst. The variety of neurological symptoms contributes to the complexity of clinical diagnosis and reinforces the need for a more specific and acceptable diagnostic approach. We demonstrated here, for the first time, that the T. multiceps DNA is present in the cerebrospinal fluid (CSF) of the infected sheep and goats. In addition, the molecular genetic marker of the mitochondrial DNA was applied phylogenetically to show that our isolates together with other T. multiceps strains comprised a monophyletic group that is a sister to Taenia krabbei. Pairwise comparison between the cox1 sequences of our study and other T. multiceps genotypes existing in the GenBank showed similarity ranging from 98 to 100%. Accordingly, the polymerase chain reaction (PCR) can be used for amplification of DNA of the parasite originated from the CSF and provides a valuable method for accurate identification of coenurosis cases.

Association of Trypanosoma vivax in extracellular sites with central nervous system lesions and changes in cerebrospinal fluid in experimentally infected goats.

Changes in cerebrospinal fluid (CSF) and anatomical and histopathological central nervous system (CNS) lesions were evaluated, and the presence of Trypanosoma vivax in CNS tissues was investigated through PCR. Twelve adult male goats were divided into three groups (G): G1, infected with T. vivax and evaluated during the acute phase; G2, infected goats evaluated during the chronic phase; and G3, consisting of non-infected goats. Each goat from G1 and G2 was infected with 1.25 × 10(5) trypomastigotes. Cerebrospinal fluid (CSF) analysis and investigation of T. vivax was performed at the 15th day post-infection (dpi) in G1 goats and on the fifth day after the manifestation of nervous system infection signs in G2 goats. All goats were necropsied, and CNS fragments from G1 and G2 goats were evaluated by PCR for the determination of T. vivax. Hyperthermia, anemia and parasitemia were observed from the fifth dpi for G1 and G2, with the highest parasitemia peak between the seventh and 21st dpi. Nervous system infection signs were observed in three G2 goats between the 30th and 35th dpi. CSF analysis revealed the presence of T. vivax for G2. Meningitis and meningoencephalitis were diagnosed in G2. PCR were positive for T. vivax in all the samples tested. In conclusion, T. vivax may reach the nervous tissue resulting in immune response from the host, which is the cause of progressive clinical and pathological manifestations of the CNS in experimentally infected goats.

Development of a diagnostic DNA probe to detect Setaria digitata: the causative parasite of cerebrospinal nematodiasis in goats, sheep and horses.

Two repetitive sequences (IpSdM and IpSdS) have been cloned and sequenced from the genome of Setaria digitata. When IpSdM (214 bp) and IpSdS (201 bp) were aligned, a high degree of homology (85%) was observed, indicating that they belong to the same family of repeats. IpSdM represents a complete repeating element while IpSdS consists of two partial repeating elements arranged in tandem. The elements are present in about 10 000 copies comprising 2.8% of the S. digitata genome. As a diagnostic probe IpSdM detects as little as 100 pg DNA of both S. digitata and S. labiato-papillosa. It can also detect a single microfilaria and a L3 larva making it a valuable tool to monitor cattle and mosquito vector populations in the prevention of cerebrospinal nematodiasis.

[Caprine arthritis-encephalitis: detection of specific antibodies in the cerebrospinal fluid and synovia]. / Caprine Arthritis-Enzephalitis: Nachweis spezifischer Antikörper in Liquor cerebrospinalis und Synovia [corrected].

Following transmission studies cerebrospinal fluid and synovia were checked for the presence of specific antibodies from ten seronegative goats derived from seropositive females and from 16 seropositive goats showing typical clinical symptoms. In the samples from the seronegative goats it was not possible to detect any specific antibodies whereas in 14 of the 16 seropositive goats specific antibodies were found in the synovia and in two goats specific antibodies could be found in the cerebrospinal fluid. The conclusion was that the local clinical symptoms may be the result of an antigen-antibody reaction.

Biochemical alterations in serum and cerebrospinal fluid in experimental acidosis in goats.

Experimental acidosis was induced in six goats aged between one and two years by administration of whole wheat grain at 100 g kg-1 bodyweight given intraruminally. Blood and cerebrospinal fluid (CSF) samples were collected from these goats before administration of wheat grain (0 hour) and thereafter at 12, 24, 48, 72, 96 and 120 hour intervals. These were analysed for serum enzyme activities and physicochemical characters of CSF. Significantly (P less than 0.05) higher activities of amylase (at 12 hours), lactate dehydrogenase (12 to 48 hours), creatine phosphokinase (12 to 48 hours), aspartate aminotransferase (12 to 24 hours), and gamma-glutamyl transferase (12 to 96 hours) were found in serum samples of acidotic goats. Changes in CSF included decrease of pH and chloride content and higher glucose values. No difference was seen in the physical character of CSF collected at different time intervals from acidotic goats.

Experimental infection of goats with Trypanosoma brucei ssp. and effects of treatment with suramin and Mel-B.

A stock of Trypanosoma brucei ssp. isolated from a naturally-infected goat in the Lambwe Valley, Kenya, induced cerebral trypanosomiasis in experimentally-infected goats. Six of nine goats with cerebral trypanosomiasis induced by this stock were cured by a single high dose of suramin (50 mg kg-1). Two other goats appeared to be cured with this dosage of suramin but later developed abnormal central nervous system (CNS) signs and parasitaemia. Parasites first appeared in the cerebrospinal fluid (CSF) and then in the blood and lymph nodes. Mel-B was also effective against primary and relapse cerebral trypanosomiasis in goats.