Marteniuk Jv

Identification of opossums (Didelphis virginiana) as the putative definitive host of Sarcocystis neurona.

Sarcocystis neurona is an apicomplexan that causes equine protozoal myeloencephalitis (EPM) in North and South America. Horses appear to be an aberrant host, because the merozoites continually divide in the central nervous system, without encysting. The natural host species has not previously been identified. The small subunit ribosomal RNA (SSURNA) gene of S. neurona was compared to those of Sarcocystis muris, Sarcocystis cruzi, Toxoplasma gondii, and Cryptosporidium parvum to identify a unique region suitable for a species-specific amplification primer. The S. neurona SSURNA primer was used in a polymerase chain reaction (PCR) assay for the purpose of identifying this organism in feces and intestinal digest of wildlife specimens. Sporocysts were isolated from 4 raccoons (Procyon lotor), 2 opossums (Didelphis virginiana), 7 skunks (Mephitis mephitis), 6 cats (Felis catus), 1 hawk (Accipiter sp.), and 1 coyote (Canis latrans). The S. neurona SSURNA PCR assay and a control PCR assay using protist-specific primers were applied to all sporocyst DNA samples. All sporocyst DNA samples tested positive on the control assay. The SSURNA PCR assay yielded a 484-bp product only when applied to opossum samples. The SSURNA gene of both opossum sporocyst samples was sequenced to determine its relationship to the S. neurona SSURNA gene. The sequence had 99.89% similarity with S. neurona. This suggests that opossums are the definitive host of S. neurona.

Risk factors for colic in the Michigan (USA) equine population

A population-based prospective epidemiological study was conducted to assess risk factors for equine colic. A stratified sample of 3925 equids in 138 randomly selected equine farms in the state of Michigan was monitored in two 12-month rounds of data collection. Incidence densities were used to describe the rate of development of colic in the study population. Mortality rates, case fatality rates and survival rates were used to describe the severity of colic on the study population. Multivariable logistic regressions with random effects (grouped according to farm) were used to identify risk factors associated with occurrence of colic. A total of 3175 equids from 132 farms from the starting population of 3925 equids in 138 farms was used in the multivariable analysis. There were 77 cases of colic reported during the study period in 62 animals. Of these animals, 54 (87%) had one case, 5 (8%) had two cases, 2 (3%) had three cases, and 1 (2%) had seven cases. Of the cases reported, 49 (64%) were non-specific diagnoses, 13 (17%) impaction/acute intestinal obstruction colics, 7 (9%) spasmodic colics, 4 (5%) sand colics, 2 (3%) gas colics, 1 (1%) verminous mesenteric arteritis, and 1 (1%) enteritis due to ingestion of moldy grain. The annual incidence density of colic in the study was 3.5 cases per 100 equid-years. The surgical treatment risk was 17% (13/77). The overall mortality risk due to colic was 0.5 deaths per 100 equids, and the case fatality risk was 13% (10/77). The case fatality risk for cases treated surgically was 31% (4/13), while the case fatality risk for non-surgical colics was 10% (7/69). Risk factors associated with significantly increased likelihood of developing colic were foaling during the study, deworming during the study, increased age, and participation in showing activities. Geldings and equids provided group drinking water from sources other than tanks, buckets and automatic waterers were significantly associated with reduced risk of colic.

Pregnancy toxemia and ketosis of ewes and does.

Pregnancy toxemia of ewes and does appears to occur when the animal cannot meet the glucose demands of the fetal-placental unit and hypoglycemia develops. There is individual variation in susceptibility, and there may be basic differences in glucose metabolism between susceptible animals and nonsusceptible animals. Increased serum NEFA and ketone body concentrations accompany the disease, but clinical signs do not appear to develop in the absence of hypoglycemia. The diagnosis is based on history, clinical signs, and the finding of ketone bodies in the urine. Numerous metabolic abnormalities develop subsequent to hypoglycemia and hyperketonemia, and these affect the prognosis. Important secondary abnormalities include acidosis, dehydration, and renal failure. Therapy is frequently unsuccessful, but frequent administration of small doses of glucose appears to be beneficial, if the other abnormalities, such as acidosis and dehydration, are controlled. Prevention can be readily achieved by nutritional means and is far more rewarding than therapy. Ewes and does must be fed in relation to their changing energy needs throughout the reproductive cycle.

Comparison of Sarcocystis neurona isolates derived from horse neural tissue

Sarcocystis neurona is a protozoan parasite that can cause neurological deficits in infected horses. The route of transmission is by fecal-oral transfer of sporocysts from opossums. However, the species identity and the lifecycle are not completely known. In this study, Sarcocystis merozoites from eight isolates obtained from Michigan horses were compared to S. neurona from a California horse (UCD1), Sarcocystis from a grackle (Cornell), and five Sarcocystis isolates from feral opossums from Michigan. Comparisons were made using several techniques. SDS-PAGE analysis with silver staining showed that Sarcocystis spp. from the eight horses appeared the same, but different from the grackle isolate. One Michigan horse isolate (MIH6) had two bands at 72 and 25kDa that were more prominent than the UCD1 isolate and other Michigan horse isolates. Western blot analysis showed that merozoites of eight of eight equine-derived isolates, and the UCD1 S. neurona isolate had similar bands when developed with serum or CSF of an infected horse. Major bands were seen at 60, 44, 30, and 16kDa. In the grackle (Cornell) isolate, bands were seen at 60, 44, 29, and 16kDa. DNA from merozoites of each of the eight equine-derived isolates and the grackle-derived isolate produced a 334bp PCR product (Tanhauser et al., 1999). Restriction fragment length polymorphism (RFLP) analysis of these horse isolates showed banding patterns characteristic for S. neurona. The grackle (Cornell) isolate had an RFLP banding pattern characteristic of other S. falcatula species. Finally, electron microscopy examining multiple merozoites of each of these eight horse isolates showed similar morphology, which differed from the grackle (Cornell) isolate. We conclude that the eight Michigan horse isolates are S. neurona species and the grackle isolate is an S. falcatula species.

Concentrations of estrone sulfate in peripheral serum of pregnant goats: Relationships with gestation length, fetal number and the occurrence of fetal death in utero

Blood sampling was initiated shortly after breeding and continued weekly through 60 to 72 days post breeding in 20 pregnant and 5 nonpregnant dairy goat does. In does not showing a return to estrus by 72 days, collection of blood was continued at 10- to 14-day intervals until 135 days post breeding. At parturition, the numbers of does delivering one, two, three, four or five kids was four, ten, three, two and one, respectively. Immunoreactive estrone sulfate (ES) was measured in serum by radioimmunoassay (RIA). In nonpregnant does, the mean+/-SD concentration of ES was 0.1+/-0.1 nmol/l (n=36 samples). At 40 to 50 days post breeding, ES concentrations increased dramatically in pregnant does. Statistically, does were classified as pregnant when serum ES increased and remained above 1.1 nmol/l (P<0.05). Serum ES in pregnant does peaked at 65 to 75 days of gestation, remained at a relative plateau through 120 days and then gradually increased for the remainder of the sampling schedule. There were positive correlations between serum ES and the number of kids born in sampling periods of 50 to 55 days and after 90 days of gestation (P<0.05). Fetal loss was documented in two does that were not included in the survey data. One doe delivered two normal kids and two mummified fetuses. Time-trend changes in ES were typical of a normal pregnancy. Another doe had a marked decrease in ES (below 0.8 nmol/l) in samples drawn on Days 94 and 108 of gestation while luteal concentrations of progesterone were maintained. This doe delivered one weak, depressed kid, one stillborn kid and a decomposed fetus at the expected time for parturition.

AN OUTBREAK OF BESNOITIOSIS IN MINIATURE DONKEYS

Fourteen miniature donkeys (Equus asinus) in a mid-Michigan herd of 38 animals presented with clinical signs of besnoitiosis, including the presence of typical tissue cysts in the ocular sclera, the buccal and nasal mucosa, together with characteristic dermatitis in specific areas of the body. The common histopathological change seen was the presence of many 100–200-μm diameter, thick walled, typical Besnoitia sp. tissue cysts together with a chronic cellular response associated with degenerating cysts. Microscopy of isolated scleral cysts and skin biopsies showed the presence of protozoal organisms consistent in morphology with that of Besnoitia bennetti bradyzoites. Molecular analysis of these parasites indicates that they differ from previously described coccidia, including Besnoitia sp., from rabbits and opossums. Isolated cases of infection with this agent have been reported infrequently in equids; however, this is the first report of an outbreak in a herd of donkeys in the United States.

A herd-level analysis of risk factors for antibodies to Sarcocystis neurona in Michigan equids.

Equine protozoal myeloencephalitis (EPM) is a neurological disease of horses and ponies caused by infection of the central nervous system with the protozoan parasite Sarcocystis neurona. A herd-level analysis of a cross-sectional study of serum antibodies to S. neurona in Michigan equids was conducted, using data collected in 1997 for study that included 1121 equids from 98 Michigan horse farms. Our objective was to identify specific herd-level risk factors associated with seropositivity. We tested associations between herd seroprevalence and various farm-management practices (including feed-storage methods and wildlife control). Multivariable models were developed for three strata based on relative opossum abundance (opossum districts). Herd seroprevalence ranged from 0 to 100% (median=57%). No risk factor was significantly associated with herd seroprevalence at P< or = 0.05 in all opossum districts. Our results suggest that equids living in areas with large opossum populations might be infected with S. neurona from multiple sources.

Descriptive epidemiology of anestrus in Michigan Holstein-Friesian cattle

Twenty-two Michigan Holstein-Friesian herds were studied to determine the incidence and epidemiology of anestrus. In 3,309 lactations studied, 42% were classified as having exhibited preservice anestrus (no estrus detected by 70 d after calving). Organic reasons (pyometra, cystic follicles, static ovaries) were identified by palpation per rectum for 237 (23%) of the cows with preservice anestrus that were examined by a veterinarian. Postservice anestrus, defined as failure to show estrus within 35 d after an unsuccessful insemination, was identified in 790 (47%) of 1,691 lactations. Veterinary examination identified 104 (20%) of postservice anestrous cows as having this condition because of organic causes. The average cow with preservice anestrus had an increase of 30 d open compared to her herdmates, and the average cow with postservice anestrus had an increase of 37 d open. Anestrous cows produced more milk than their unaffected herdmates in both the current and the previous lactation. Analysis of composite lactation curves indicates that, for preservice anestrus, this additional milk production was obtained gradually over the entire lactation. Anestrous cows were culled at a significantly higher rate than their herdmates. Cows with preservice anestrus were more likely to have begun their lactation in the spring months.

Association between hyperinsulinaemia and laminitis severity at the time of pituitary pars intermedia dysfunction diagnosis

BACKGROUND Hyperinsulinaemia is the suspected component of insulin dysregulation having the strongest association with laminitis and occurs variably in equids with pituitary pars intermedia dysfunction (PPID). OBJECTIVES We hypothesised that magnitude of hyperinsulinaemia correlates with laminitis severity in PPID-affected equids. Furthermore, we hypothesised that owners can be unaware of chronic endocrinopathic laminitis. STUDY DESIGN Cross-sectional study. METHODS Serum insulin concentrations, owner-reported laminitis history and radiographic evidence of laminitis were determined in 38 client-owned horses and ponies with confirmed PPID. Laminitis severity was classified into four categories (normal [nonlaminitic], mild, moderate or severe laminitis) based on degree of distal phalangeal rotation. Animals were also categorised as normoinsulinaemic (<20 μU/ml), mildly hyperinsulinaemic (20-50 μU/ml) and severely hyperinsulinaemic (>50 μU/ml). One-way ANOVA, t tests and Fishers exact tests were performed. RESULTS While owners reported laminitis in 37% of animals, 76% were laminitic based on study criteria (P = 0.01). Owners reported laminitis more frequently in hyperinsulinaemic vs. normoinsulinaemic animals; recognition increased with severity of hyperinsulinaemia (P = 0.03). Mean insulin concentrations were higher in equids with moderate to severe radiographic laminitis (geometric mean 74.1, 95% confidence interval (CI) 38.4-143.1 uU/ml) vs. those classified radiographically as normal to mild (31.9, 95% CI 21.1-48.1 uU/ml P = 0.03). MAIN LIMITATIONS Dynamic insulin testing was not performed; some normoinsulinaemic animals might have had subtle insulin dysregulation. CONCLUSIONS Although radiographic abnormalities were present in most animals at the time of PPID diagnosis, chronic laminitis remained unrecognised by many owners. Owner awareness of laminitis increased with severity of hyperinsulinaemia and higher insulin concentrations were detected in association with more severe radiographic changes. The Summary is available in Chinese - See Supporting Information.

Bacterial and fungal microflora on the external genitalia of male donkeys (Equus asinus)

This study was undertaken to investigate the bacterial and fungal microflora on the external genitalia of a population of healthy male donkeys in the state of Michigan, USA. The aim was to identify and determine the frequency of occurrence of these microorganisms using seven different isolation media and standard microbiological procedures. The sites (urethral fossa [fossa glandis], dorsal diverticulum of the urethral sinus, distal urethra, and penile surface) in the distal reproductive tract were cultured and each isolated microorganism identified. Ten different genera of gram-positive bacteria, eight different genera of gram-negative bacteria, and two genera of fungi were isolated from the external genitalia of the 43 donkeys in this study. All 43 donkeys yielded gram-positive bacteria (2-8 species) from all four sites sampled. Arcanobacterium spp., Corynebacterium spp., and Bacillus spp. were the most frequently isolated gram-positive bacteria. Gram-negative bacteria were cultured from 16 (37.2%) of the 43 donkeys, with Acinetobacterlwoffii (16.3%), Oligella urethralis (11.6%), and Taylorellaasinigenitalis (9.3%), the most frequently isolated. Fungi were cultured from only 5 (11.6%) of the 43 donkeys, with Rhizopus spp. isolated from 3 (7.0%) and Cladosporium spp. from 2 (4.7%) individuals. The testes and epididymides collected from 40 donkeys at time of castration were culture negative. Few differences were found in the bacterial flora between prepubertal and mature intact and castrated donkeys. Of notable interest was the scarcity of known equine pathogens across the population tested and isolation of T. asinigenitalis from normal donkeys, especially prepubertal individuals and previously castrated males.