Neuronal ceroid lipofuscinosis: clinical and morphologic findings in nine affected Polish Owczarek Nizinny (PON) dogs

OBJECTIVE: The aim of this study was to characterize the clinical and morphologic features of neuronal ceroid lipofuscinosis (NCL) in the Polish Owczarek Nizinny (PON) breed of dog. ANIMALS: Nine Swedish PON dogs of both sexes were included in the study. PROCEDURE: All dogs underwent a detailed clinical evaluation, with emphasis on ophthalmic exams. Histopathology and electron microscopy were performed on the eyes, brain and various internal organs. Immunohistochemical staining for detection of sphingolipid activator proteins (SAPs) and mitochondrial ATP synthase (SCMAS) was performed on the eyes and brain. RESULTS: The dogs showed behavioral abnormalities, motor disturbances and visual impairment or blindness. Pupillary responses were abnormal while fundus changes varied from normal to severe retinal atrophy. Electroretinography (ERG) showed variable changes, from slight alterations in the process of dark adaptation to severely reduced or nonrecordable ERG a- and b-wave amplitudes. Histopathology revealed intracytoplasmic storage bodies within neurons of the brain and in retinal cells, especially the retinal pigment epithelium (RPE). Round to oval granular type of inclusion bodies, known as granular osmiophilic dense deposits (GRODS), were found in neuronal cells in the brain and in the retina. Immunohistochemistry identified the storage material in the brain and retina as consisting of SAPs. CONCLUSION: The presently described NCL disease in PON dogs shows similarities to previously recorded cases in the Miniature Schnauzer. The closest human equivalent to this disease is infantile NCL (CLN1), in which the major stored proteins are SAPs and the ultrastructure of the inclusion bodies of neuronal cells is granular.     

Co-infection with Mycoplasma haemofelis and 'Candidatus Mycoplasma haemominutum' in three cats from Brazil

The two most common haemotropic Mycoplasma of cats, Mycoplasma haemofelis and 'Candidatus Mycoplasma haemominutum' have been identified using molecular techniques in all continents, except Antarctica. We report the first molecular characterization in South America of a dual infection with M haemofelis and 'Candidatus Mycoplasma haemominutum' in three domestic cats. The 16S ribosomal RNA gene was amplified in three anaemic cats in which haemoplasma organisms were seen attached to the erythrocytes in the peripheral blood smear. Bands of the expected size for M haemofelis and 'Candidatus Mycoplasma haemominutum' were observed in all three cats. The 393 bp segment of one of the amplicons had a similarity value of 100% to M haemofelis, whereas the other amplicon, a 192 bp segment, was 100% similar to 'Candidatus Mycoplasma haemominutum'. After diagnosis, two cats received blood transfusion and they were all treated with doxycycline. All three cats recovered uneventfully.     

Importance of prudent antibiotic use on dairy farms in South Carolina: a pilot project on farmers' knowledge, attitudes and practices

Inappropriate use of antibiotics in humans and animals contributes to decreased antimicrobial susceptibility in bacteria of medical importance. Resistant bacteria being transferred from animals to humans are causing public health concern. In-person interviews were conducted with 20 dairy farmers in rural counties of South Carolina to determine farmers' knowledge and attitudes about prudent antibiotic use among livestock. Four focus groups (n = 22) were also conducted to ascertain farmers' specific information needs about proper antibiotic use. Survey results showed that participants (100%) typically determined a need for antibiotic treatment using symptom assessment and reported following some form of operating procedures regarding administration of antibiotics. Few farmers (32%) had actual written antibiotic protocols. Preferred information sources about antibiotics were veterinarians (100%) and other dairy farmers (50%). Most farmers (86%) were not concerned that overuse of antibiotics in animals could result in antibiotic resistance among farm workers. Qualitative analysis of focus groups revealed significant barriers to following proper antibiotic procedures including limited finances and lack of time. The need for bilingual educational resources for Hispanic/Latino dairy workers was expressed. Desired formats for educational materials were posters, flowcharts, videos, and seminars. Education of South Carolina dairy farmers by veterinarians and public health professionals on the appropriate use of antibiotics in dairy cattle is needed to ensure antibiotic effectiveness in both animals and humans.     

Comparison of duplex PCR and microscopic techniques for the identification of Babesia bigemina and Babesia bovis in engorged female ticks of Boophilus microplus

A single-step duplex polymerase chain reaction (PCR) technique and traditional microscopic examination of haemolymph smears were used to detect Babesia bigemina and/or Babesia bovis infection in engorged female ticks of Boophilus microplus recovered from calves raised in an endemic area of the State of Minas Gerais, Brazil. In the PCR amplification of tick-derived DNA, pairs of oligonucleotide primers specific for a 278-bp sequence from B. bigemina and for a 350-bp sequence from B. bovis were used conjointly. The microscopic examination of haemolymph revealed that 16.7% of the engorged ticks were infected with Babesia spp., although no significant differences (rho > 0.05) were found in the infection rate of ticks collected from calves of different age groups. PCR analysis showed that 77.8% of the engorged ticks whose haemolymph contained sporokinetes were infected with B. bigemina, 7.8% with B. bovis and 14.4% with both protozoan species. However, the PCR assay further revealed that, amongst the engorged female ticks whose haemolymph was apparently negative for the presence of sporokinetes, 15.6% were infected with B. bigemina, 2.2% with B. bovis and 10.0% with both species. The duplex PCR method is thus more efficient and sensitive than the microscopic assay and also permits facile identification of the protozoa species present in engorged female ticks.     

Application of an electrometric method for measurement of blood cholinesterase activities in sheep, goats and cattle treated with organophosphate insecticides

A modified electrometric cholinesterase method has been described for use in ruminants exposed to organophosphate insecticides. The method was used to measure cholinesterase activities in the plasma and erythrocytes of sheep, goats and cattle treated with organophosphate insecticides under field conditions. The animals were treated topically to control ectoparasites with malathion (0.05%) or diazinon (0.06%). The treated animals did not suffer from adverse effects related to cholinesterase inhibition. However, 2 h after treatments, plasma and erythrocyte cholinesterase activities significantly decreased in sheep by 20% and 39% and in goats by 18% and 30%, respectively, when compared with respective untreated control values. Diazinon did not significantly affect plasma and erythrocyte cholinesterase activities in the cattle. The results suggest that the described electrometric method could be efficiently used for detecting cholinesterase inhibition in ruminants exposed to organophosphate insecticides.     

Incidence and effects of West Nile virus infection in vaccinated and unvaccinated horses in California

A prospective cohort study was used to estimate the incidence of West Nile virus (WNV) infection in a group of unvaccinated horses (n = 37) in California and compare the effects of natural WNV infection in these unvaccinated horses to a group of co-mingled vaccinated horses (n = 155). Horses initially were vaccinated with either inactivated whole virus (n = 87) or canarypox recombinant (n = 68) WNV vaccines during 2003 or 2004, prior to emergence of WNV in the region. Unvaccinated horses were serologically tested for antibodies to WNV by microsphere immunoassay incorporating recombinant WNV E protein (rE MIA) in December 2003, December 2004, and every two months thereafter until November 2005. Clinical neurologic disease attributable to WNV infection (West Nile disease (WND)) developed in 2 (5.4%) of 37 unvaccinated horses and in 0 of 155 vaccinated horses. One affected horse died. Twenty one (67.7%) of 31 unvaccinated horses that were seronegative to WNV in December, 2004 seroconverted to WNV before the end of the study in November, 2005. Findings from the study indicate that currently-available commercial vaccines are effective in preventing WND and their use is financially justified because clinical disease only occurred in unvaccinated horses and the mean cost of each clinical case of WND was approximately 45 times the cost of a 2-dose WNV vaccination program.     

IgE-bearing cells in bronchoalveolar lavage fluid and allergen-specific IgE levels in sera from RAO-affected horses

Recurrent airway obstruction (RAO) is a common condition in stabled horses characterized by small airway inflammation, airway neutrophilia and obstruction following exposure of susceptible horses to mouldy hay and straw and is thus regarded as a hypersensitivity reaction to mould spores. However, the role of immunoglobulin E antibodies (IgE) in the pathogenesis of RAO is unclear. We hypothesized that the number of cells with receptor-bound IgE in bronchoalveolar lavage fluid (BALF) and IgE levels in serum would be higher in RAO-affected than in healthy horses living in the same environment. Therefore, IgE-positive (+) cells were identified by immunocytochemistry on cytospins from BALF and counted. IgE levels against the mould extracts Aspergillus fumigatus (Asp. f.) and Alternaria alternata (Alt. a.) and the recombinant mould allergen Aspergillus fumigatus 8 (rAsp f 8) were measured by enzyme-linked immunosorbent assay (ELISA) in the sera of seven RAO-affected and 22 clinically healthy mature horses housed in the same conventional stable environment. After correcting for the number of neutrophils, there were no significant differences in IgE+ cells on cytospins from BALF between both groups of horses (5% versus 7%, P > 0.1). Serum IgE levels against the mould extracts were significantly higher in RAO-affected than in clinically healthy horses [median = 119 versus 66 relative ELISA units (REU), P < 0.05]. Furthermore, significantly more RAO-affected than healthy horses had detectable serum IgE against the recombinant allergen rAsp f 8 (4/7 and 3/22, respectively, P < 0.05). Age had no significant effect on BALF cell ratios or on specific serum IgE levels. These results show that high IgE levels against mould antigens are associated with RAO under controlled environmental conditions but ranges of mould-specific serum IgE levels overlapped too much between diseased and clinically healthy animals to be of any diagnostic value. Further studies are needed to assess whether IgE-mediated reactions contribute to the pathogenesis of RAO.     

Synovial fluid analysis and bacterial findings in arthritic joints of juvenile male camel (Camelus dromedarius) calves

Eighteen synovial fluid samples from 11 male dromedarian calves, 9-12 month old, were analysed cytologically and bacteriologically. Calves were lame and all joints were grossly swollen. The mean +/- SD of total nucleated cell count was 7970 +/- 5000 cells/microl (range 2800-20,000 cells/microl). Polymorphonuclear (PMN) leucocytes were the predominant cell type. The mean +/- SD of absolute and percentages of each cell type were as follows: PMN leucocytes 5518 +/- 3600 cells/microl and 68 +/- 19%, monocytes/macrophages 1600 +/- 1120 cells/microl and 26 +/- 17%, lymphocytes 830 +/- 140 cells/microl and 8 +/- 7%, and red blood cell 350 +/- 130 cells/microl. The mean +/- SD of total protein concentration was 3.5 +/- 1 g/dl (range 2.5-5 g/dl). The most commonly isolated bacteria were non-haemolytic streptococci spp., followed by Arcanobacterium pyogenes and Staphylococcus aureus. No bacterial growth was obtained in eight samples and non-revealed Mycoplasma spp.     

Phylogenetic characterization of rabies virus isolates from Carnivora in Brazil

The incidence of canine rabies has been widely reported in Brazil, and new rabies virus (RV) variants, genetically similar to canine RV, have recently been isolated from foxes. In order to derive the epidemiological characteristics of Brazilian Carnivora RV, Brazilian RVs isolated from dogs, cats, and foxes were genetically analyzed. Brazilian Carnivora RV isolates were divided into 2 main lineages. The predominant lineage was found in dogs and cats, which included the Argentinean and Bolivian Carnivora RV isolates, and was extensively distributed throughout Brazil and surrounding countries. The other lineage consisted of three sublineages containing Brazilian dog and fox RV isolates, with the dog sublineages located on an internal branch of 2 fox sublineages, suggesting that RV transmission events might have occurred between foxes and dogs in the past. These results suggest that contact between dogs and wildlife has the potential to generate new rabies variants and that it is important to control RV infection cycles in both dogs and wildlife to prevent spread of rabies infection.     

Milk yield, age at first calving, and the risk of BSE: an analysis at the farm level in France

The epidemic of bovine spongiform encephalopathy (BSE) in France, as in the UK, has affected dairy cattle much more than beef cattle. However, the intensification of dairy herd management as a risk factor for BSE has not to date been analyzed. For this purpose, two databases were merged: the French Milk Records database, and the French BSE database, which can be considered as being devoid of notification bias since July 2001, when systematic tests were implemented. Only pure Holstein herds were considered, which represent the vast majority of total and BSE-affected dairy herds in France. A case-control study was designed so that 20 control herds were matched to each case herd according to the location of the farm and the year of birth of the index case. Three thousand and forty five farms were included, among which 145 with a BSE case notified between July 2001 and July 2003, and 2900 controls. With respect to the risk of BSE, odds ratios for each class of milk yield and age at first calving were estimated by using conditional logistic regression models with appropriate adjustments to herd size. The two main results were the following: firstly, whereas most Holstein herds, with average production between 7000 and 10,000kg, had nearly the same BSE risk, a small category of very intensive herds, with annual milk yields above 10,000kg, were significantly more at risk than the other herds. Secondly, a very early first calving (under 26 months of age) was found to be at risk for BSE as compared to other categories, independently of the milk yield. These results are discussed in the light of the known age-dependent susceptibility to BSE.